<?xml version="1.0" encoding="UTF-8"?>
<!DOCTYPE article PUBLIC "-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" "journalpublishing.dtd">
<article article-type="research-article" dtd-version="2.3" xml:lang="EN" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">
<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Cell Dev. Biol.</journal-id>
<journal-title>Frontiers in Cell and Developmental Biology</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Cell Dev. Biol.</abbrev-journal-title>
<issn pub-type="epub">2296-634X</issn>
<publisher>
<publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="publisher-id">1197109</article-id>
<article-id pub-id-type="doi">10.3389/fcell.2023.1197109</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Cell and Developmental Biology</subject>
<subj-group>
<subject>Original Research</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group>
<article-title>Sin3a associated protein 130&#xa0;kDa, sap130, plays an evolutionary conserved role in zebrafish heart development</article-title>
<alt-title alt-title-type="left-running-head">DeMoya et al.</alt-title>
<alt-title alt-title-type="right-running-head">
<ext-link ext-link-type="uri" xlink:href="https://doi.org/10.3389/fcell.2023.1197109">10.3389/fcell.2023.1197109</ext-link>
</alt-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>DeMoya</surname>
<given-names>Ricardo A.</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/2416956/overview"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Forman-Rubinsky</surname>
<given-names>Rachel E.</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/2341421/overview"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Fontaine</surname>
<given-names>Deon</given-names>
<suffix>Jr</suffix>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/2265339/overview"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Shin</surname>
<given-names>Joseph</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/2417102/overview"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Watkins</surname>
<given-names>Simon C.</given-names>
</name>
<xref ref-type="aff" rid="aff2">
<sup>2</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/40071/overview"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Lo</surname>
<given-names>Cecilia W.</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<uri xlink:href="https://loop.frontiersin.org/people/150153/overview"/>
</contrib>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>Tsang</surname>
<given-names>Michael</given-names>
</name>
<xref ref-type="aff" rid="aff1">
<sup>1</sup>
</xref>
<xref ref-type="corresp" rid="c001">&#x2a;</xref>
<uri xlink:href="https://loop.frontiersin.org/people/38690/overview"/>
</contrib>
</contrib-group>
<aff id="aff1">
<sup>1</sup>
<institution>Department of Developmental Biology</institution>, <institution>University of Pittsburgh School of Medicine</institution>, <addr-line>Pittsburgh</addr-line>, <addr-line>PA</addr-line>, <country>United States</country>
</aff>
<aff id="aff2">
<sup>2</sup>
<institution>Department of Cell Biology</institution>, <institution>University of Pittsburgh School of Medicine</institution>, <addr-line>Pittsburgh</addr-line>, <addr-line>PA</addr-line>, <country>United States</country>
</aff>
<author-notes>
<fn fn-type="edited-by">
<p>
<bold>Edited by:</bold> <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/45550/overview">Diego Franco</ext-link>, University of Ja&#xe9;n, Spain</p>
</fn>
<fn fn-type="edited-by">
<p>
<bold>Reviewed by:</bold> <ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/1163770/overview">Jiandong Liu</ext-link>, University of North Carolina at Chapel Hill, United States</p>
<p>
<ext-link ext-link-type="uri" xlink:href="https://loop.frontiersin.org/people/1125580/overview">Marina Campione</ext-link>, National Research Council (CNR), Italy</p>
</fn>
<corresp id="c001">&#x2a;Correspondence: Michael Tsang, <email>tsang@pitt.edu</email>
</corresp>
</author-notes>
<pub-date pub-type="epub">
<day>30</day>
<month>08</month>
<year>2023</year>
</pub-date>
<pub-date pub-type="collection">
<year>2023</year>
</pub-date>
<volume>11</volume>
<elocation-id>1197109</elocation-id>
<history>
<date date-type="received">
<day>30</day>
<month>03</month>
<year>2023</year>
</date>
<date date-type="accepted">
<day>17</day>
<month>08</month>
<year>2023</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#xa9; 2023 DeMoya, Forman-Rubinsky, Fontaine, Shin, Watkins, Lo and Tsang.</copyright-statement>
<copyright-year>2023</copyright-year>
<copyright-holder>DeMoya, Forman-Rubinsky, Fontaine, Shin, Watkins, Lo and Tsang</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<p>This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</p>
</license>
</permissions>
<abstract>
<p>Hypoplastic left heart syndrome (HLHS) is a congenital heart disease where the left ventricle is reduced in size. A forward genetic screen in mice identified SIN3A associated protein 130&#xa0;kDa (<italic>Sap130</italic>), part of the chromatin modifying SIN3A/HDAC complex, as a gene contributing to the etiology of HLHS. Here, we report the role of zebrafish <italic>sap130</italic> genes in heart development. Loss of <italic>sap130a,</italic> one of two <italic>Sap130</italic> orthologs, resulted in smaller ventricle size, a phenotype reminiscent to the hypoplastic left ventricle in mice. While cardiac progenitors were normal during somitogenesis, diminution of the ventricle size suggest the Second Heart Field (SHF) was the source of the defect. To explore the role of <italic>sap130a</italic> in gene regulation, transcriptome profiling was performed after the heart tube formation to identify candidate pathways and genes responsible for the small ventricle phenotype. Genes involved in cardiac differentiation and cardiac function were dysregulated in <italic>sap130a</italic>, but not in <italic>sap130b</italic> mutants. Confocal light sheet analysis measured deficits in cardiac output in <italic>MZsap130a</italic> supporting the notion that cardiomyocyte maturation was disrupted. Lineage tracing experiments revealed a significant reduction of SHF cells in the ventricle that resulted in increased outflow tract size. These data suggest that <italic>sap130a</italic> is involved in cardiogenesis via regulating the accretion of SHF cells to the growing ventricle and in their subsequent maturation for cardiac function. Further, genetic studies revealed an interaction between <italic>hdac1</italic> and <italic>sap130a</italic>, in the incidence of small ventricles. These studies highlight the conserved role of Sap130a and Hdac1 in zebrafish cardiogenesis.</p>
</abstract>
<kwd-group>
<kwd>cardiac development</kwd>
<kwd>second heart field</kwd>
<kwd>SIN3A/HDAC complex</kwd>
<kwd>congenital heart disease</kwd>
<kwd>zebrafish</kwd>
</kwd-group>
<contract-num rid="cn001">R01HL142788</contract-num>
<contract-sponsor id="cn001">National Institutes of Health<named-content content-type="fundref-id">10.13039/100000002</named-content>
</contract-sponsor>
<custom-meta-wrap>
<custom-meta>
<meta-name>section-at-acceptance</meta-name>
<meta-value>Molecular and Cellular Pathology</meta-value>
</custom-meta>
</custom-meta-wrap>
</article-meta>
</front>
<body>
<sec sec-type="intro" id="s1">
<title>Introduction</title>
<p>Congenital heart diseases (CHDs) affect approximately 1% of live births per year and causes have been attributed to environmental and genetic factors (<xref ref-type="bibr" rid="B66">Nora, 1968</xref>; <xref ref-type="bibr" rid="B23">Fahed et al., 2013</xref>; <xref ref-type="bibr" rid="B18">Costain et al., 2016</xref>). Hypoplastic left heart syndrome (HLHS) is a critical CHD characterized by a reduced volume in the left ventricle and aortic and valve malformations (<xref ref-type="bibr" rid="B17">Connor and Thiagarajan, 2007</xref>; <xref ref-type="bibr" rid="B5">Barron et al., 2009</xref>). The genetic etiology of HLHS is complex and genetically heterogenous. Mouse models of HLHS were recovered from a large-scale mutagenesis screen (<xref ref-type="bibr" rid="B57">Liu et al., 2017</xref>), and among 8 lines, the <italic>Ohia</italic> mutant line was identified to have a digenic etiology for HLHS. This is comprised of mutations in SIN3A associated protein 130&#xa0;kDa (SAP130) and protocadherin 9 (PCDHA9) that together causes HLHS comprising hypoplasia of all left-sided heart structures including the ventricle, aorta/aortic valve, and mitral valve. In pigs a CRISPR generated SAP130 allele caused embryonic lethality and tricuspid dysplasia and atresia, indicating SAP130 involvement in cardiac development in higher vertebrates (<xref ref-type="bibr" rid="B26">Gabriel et al., 2021</xref>). In zebrafish, maternal zygotic <italic>sap130a</italic> (<italic>MZsap130a</italic>) mutants resulted in a diminutive ventricle by 72&#xa0;h post fertilization (hpf), confirming that SAP130 retains a conserved function among vertebrates during heart development (<xref ref-type="bibr" rid="B57">Liu et al., 2017</xref>; <xref ref-type="bibr" rid="B26">Gabriel et al., 2021</xref>).</p>
<p>SAP130 was identified as an interacting protein in the SIN3A complex, binding both SIN3A and Histone Deacetylase 1 (HDAC1), thought to stabilize the complex. It was theorized that the SAP130 C-terminus functioned as a transcriptional repressor in association with the SIN3A complex, while the N-terminus paradoxically could function as an activator (<xref ref-type="bibr" rid="B25">Fleischer et al., 2003</xref>). A knock-out allele of SAP130 in mice is peri-implantation lethal, unlike global knockouts of HDAC1 and SIN3A which die at later stages of development (<xref ref-type="bibr" rid="B51">Lagger et al., 2002</xref>; <xref ref-type="bibr" rid="B19">Dannenberg et al., 2005</xref>; <xref ref-type="bibr" rid="B57">Liu et al., 2017</xref>). These suggest multiple roles and stages of development where SAP130/SIN3A/HDAC1 are critical for life. SIN3A and HDACs epigenetically regulate transcription through histone and non-histone deacetylation events and are classically associated with gene repression. However, some studies have shown this complex to be a transcriptional activator in other contexts (<xref ref-type="bibr" rid="B33">Han et al., 2011</xref>; <xref ref-type="bibr" rid="B48">Kadamb et al., 2013</xref>; <xref ref-type="bibr" rid="B1">Adams et al., 2018</xref>). HDACs have been reported to regulate many aspects of development, including cardiac development in zebrafish, mouse, and chick models, as evidenced by treatment with a pan HDAC small molecule inhibitor, Trichostatin A (<xref ref-type="bibr" rid="B35">Hargreaves and Crabtree, 2011</xref>; <xref ref-type="bibr" rid="B59">McKinsey, 2011</xref>; <xref ref-type="bibr" rid="B58">Martinez et al., 2015</xref>). Zebrafish studies have revealed that <italic>hdac1</italic> is involved in Second Heart Field (SHF) development and in adult cardiac regeneration (<xref ref-type="bibr" rid="B78">Song et al., 2019</xref>; <xref ref-type="bibr" rid="B11">Buhler et al., 2021</xref>). In zebrafish, <italic>hdac1</italic> mutants have less cardiomyocytes (CMs) in the ventricle while inhibition of <italic>hdac1</italic> (and other class I HDACs) reveal reduced proliferation during regenerative events (<xref ref-type="bibr" rid="B61">Montgomery et al., 2007</xref>; <xref ref-type="bibr" rid="B65">Nambiar et al., 2007</xref>; <xref ref-type="bibr" rid="B78">Song et al., 2019</xref>; <xref ref-type="bibr" rid="B11">Buhler et al., 2021</xref>). Zebrafish <italic>hdac1</italic> mutants are embryonic lethal, similar to the mouse models, but <italic>MZsap130a</italic> mutants are viable as adults suggesting that <italic>hdac1</italic> and <italic>sap130a</italic> may have distinct functions in zebrafish cardiogenesis.</p>
<p>In addition to the Sin3a/Hdac1 complex, related chromatin modifying complexes like the BAF complex, have been shown to be involved in cardiogenesis (<xref ref-type="bibr" rid="B55">Lickert et al., 2004</xref>; <xref ref-type="bibr" rid="B87">Wang et al., 2004</xref>; <xref ref-type="bibr" rid="B79">Stankunas et al., 2008</xref>; <xref ref-type="bibr" rid="B34">Hang et al., 2010</xref>; <xref ref-type="bibr" rid="B35">Hargreaves and Crabtree, 2011</xref>; <xref ref-type="bibr" rid="B81">Takeuchi et al., 2011</xref>; <xref ref-type="bibr" rid="B53">Lei et al., 2012</xref>; <xref ref-type="bibr" rid="B77">Singh and Archer, 2014</xref>; <xref ref-type="bibr" rid="B64">Nakamura et al., 2016</xref>; <xref ref-type="bibr" rid="B89">Xiao et al., 2016</xref>; <xref ref-type="bibr" rid="B80">Sun et al., 2018</xref>; <xref ref-type="bibr" rid="B2">Alfert et al., 2019</xref>; <xref ref-type="bibr" rid="B37">Hota et al., 2019</xref>; <xref ref-type="bibr" rid="B54">Lei et al., 2019</xref>; <xref ref-type="bibr" rid="B14">Chen et al., 2022</xref>; <xref ref-type="bibr" rid="B4">Auman et al., 2023</xref>). A study describing the loss of <italic>smarcc1a</italic>, a BAF chromatin remodeling complex protein, in zebrafish resulted in dysmorphic cardiac chambers further highlighting the importance of chromatin remodeling in proper heart formation (<xref ref-type="bibr" rid="B4">Auman et al., 2023</xref>). Another part of the BAF complex in zebrafish <italic>brg1</italic>, when mutated reveals a reduction in CM proliferation leading to a smaller ventricle after 28hpf. The <italic>brg1</italic> mutants reveal changes in a working myocardium marker <italic>nppa</italic>, similar to mouse Brg1 mutants (<xref ref-type="bibr" rid="B81">Takeuchi et al., 2011</xref>). Other types of epigenetic regulation such as methylation are shown to be paired with chromatin remodeling events and are involved in cardiogenic processes (<xref ref-type="bibr" rid="B12">Carrozza et al., 2005</xref>; <xref ref-type="bibr" rid="B47">Joshi and Struhl, 2005</xref>; <xref ref-type="bibr" rid="B49">Keogh et al., 2005</xref>; <xref ref-type="bibr" rid="B10">Brown et al., 2006</xref>; <xref ref-type="bibr" rid="B22">Donlin et al., 2012</xref>; <xref ref-type="bibr" rid="B86">Voelkel et al., 2013</xref>; <xref ref-type="bibr" rid="B77">Singh and Archer, 2014</xref>; <xref ref-type="bibr" rid="B90">Xiao et al., 2018</xref>; <xref ref-type="bibr" rid="B94">Zhu et al., 2018</xref>; <xref ref-type="bibr" rid="B7">Bisserier et al., 2021</xref>). SET and MYND domain-containing lysine methyltransferase 4 (<italic>smyd4</italic>) mutants also result in reduced ventricle size in zebrafish and mouse, suggesting there is a common requirement of gene regulation for specifying heart organ size in vertebrates (<xref ref-type="bibr" rid="B82">Trotter and Archer, 2008</xref>). RNA sequencing (RNA-seq) analysis of <italic>smyd4</italic> zebrafish mutants revealed dysregulation of cardiac muscle contraction and metabolism genes. Moreover, cell culture studies revealed human SMYD4 and HDAC1 interact, further supporting a central requirement for <italic>hdac1</italic> in zebrafish cardiogenesis (<xref ref-type="bibr" rid="B90">Xiao et al., 2018</xref>). Taken together these suggest a potential epigenetic role for <italic>sap130a</italic> during development as part of the Sin3a complex.</p>
<p>Here we investigate the role of <italic>sap130</italic> genes in zebrafish by studying mutations in both <italic>sap130a</italic> and <italic>sap130b</italic>. Transcriptome profiling of 36hpf <italic>MZsap130a</italic> mutants revealed over 5,000 genes to be differentially expressed, including genes involved in the cardiac development and function. In genetic studies, an increase in embryos with small ventricles (SVs) were noted in <italic>MZsap130a</italic> embryos that were also heterozygous for <italic>hdac1</italic>. Furthermore, <italic>MZsin3ab</italic> mutants exhibit a SV phenotype at 48hpf. Collectively, these studies suggest a role for <italic>sin3ab</italic>/<italic>hdac1</italic>/<italic>sap130a</italic> in the SHF during zebrafish cardiogenesis.</p>
</sec>
<sec sec-type="materials|methods" id="s2">
<title>Materials and methods</title>
<sec id="s2-1">
<title>Zebrafish husbandry</title>
<p>All zebrafish experiments and protocols were performed according to protocols approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Pittsburgh in agreement with NIH guidelines. Wild-type AB&#x2a;, <italic>Tg</italic>(<italic>myl7:GFP</italic>)<sup>
<italic>twu34</italic>
</sup> (<xref ref-type="bibr" rid="B38">Huang et al., 2003</xref>), <italic>Tg</italic>(<italic>nkx2.5:kaeda</italic>)<sup>
<italic>fb9</italic>
</sup> (<xref ref-type="bibr" rid="B31">Guner-Ataman et al., 2013</xref>), <italic>sap130a</italic>
<sup>
<italic>pt32a</italic>
</sup> (<xref ref-type="bibr" rid="B57">Liu et al., 2017</xref>), <italic>hdac1</italic>
<sup>
<italic>b382</italic>
</sup> (<xref ref-type="bibr" rid="B41">Ignatius et al., 2013</xref>).</p>
<p>Adult tail fin clips or whole embryos for genotyping assays was performed as previously described (<xref ref-type="bibr" rid="B45">Jing, 2012</xref>). Restriction fragment length polymorphism (RFLP) genotyping for <italic>sap130a</italic>
<sup>
<italic>pt32a</italic>
</sup>, <italic>sap130b</italic>
<sup>
<italic>pt35b</italic>
</sup>, <italic>sin3ab</italic>
<sup>
<italic>pt36a</italic>
</sup> and <italic>hdac1</italic>
<sup>
<italic>b382</italic>
</sup> used the primers and enzymes listed in <xref ref-type="sec" rid="s11">Supplementary Table S1</xref>.</p>
</sec>
<sec id="s2-2">
<title>CRISPR/Cas9 mutant allele generation</title>
<p>The CRISPR/Cas9 protocol (<xref ref-type="bibr" rid="B27">Gagnon et al., 2014</xref>) was used to establish mutant lines. This protocol used Sp6 <italic>in vitro</italic> transcribed sgRNAs targeting the sequence ccg&#x200b;TGG&#x200b;GAG&#x200b;GGA&#x200b;AAA&#x200b;CAA&#x200b;TGC&#x200b;TG for <italic>sap130b</italic> and cct&#x200b;GCT&#x200b;CCT&#x200b;CTT&#x200b;CAG&#x200b;CCA&#x200b;TAC&#x200b;AG for <italic>sin3ab</italic>, where lower case letters represent the protospacer motif sequence. sgRNA was incubated at room temperature with Cas9 protein (NEB, Cat&#x23; M0646T). AB&#x2a; embryos were injected at the one-cell stage with the sgRNA and Cas9 cocktail in a 1&#xa0;nL volume at 25&#xa0;pg sgRNA/nL. RFLP was performed to determine protected mutated bands present 24hrs after injection to determine gRNA efficiency and injected embryos were raised to adults outcrossed to AB&#x2a;. DNA mutations in <italic>sap130b</italic> and <italic>sin3ab</italic> were verified by PCR TOPO-TA cloning (ThermoFisher, &#x23;K4575J10) from adult heterozygous animals and Sanger sequenced. gRNA sequence information <xref ref-type="sec" rid="s11">Supplementary Table S1</xref>.</p>
</sec>
<sec id="s2-3">
<title>Imaging</title>
<p>A Leica M205 FA stereomicroscope was used to take images of the hearts from <italic>Tg(myl7:EGFP)</italic> WT and mutant embryos at 36 and 48hpf. For imaging the <italic>Tg(myl7:memGFP)</italic> OFT, a Nikon A1 inverted confocal microscope was used at 72hpf. <italic>Tg(myl7:memGFP)</italic> embryos were anesthetized in 7x MS-222/10&#xa0;mM BDM (2,3-butanedione monoxime) and mounted in low melting agarose on MaTek glass bottom petri dish (MaTek, Part No: P35G-1.5&#x2013;14-C) and imaged at a 40x water immersion. For counting cardiomyocytes at 72hpf, <italic>Tg (myl7:memGFP)</italic> and <italic>MZsap130a;Tg(myl7:EGFP)</italic> were injected with 50&#xa0;pg of <italic>H2b-mCherry</italic> mRNA at the 1-2 cell stage. Injected embryos were incubated at 28&#xb0;C and mounted on an inverted confocal microscope at 72hpf on a Nikon A1 microscope. Ventricular cardiomyocytes were designated as positive for both <italic>mCherry</italic> nuclei and membrane GFP expression using Fiji ImageJ and the orthogonal views tool.</p>
</sec>
<sec id="s2-4">
<title>ConSurf and R generated phylogenetic trees and protein diagram</title>
<p>ConSurf (<ext-link ext-link-type="uri" xlink:href="https://consurf.tau.ac.il/consurf_index.php">https://consurf.tau.ac.il/consurf_index.php</ext-link>) was used to align multiple Sap130 protein sequences across many species (<xref ref-type="bibr" rid="B6">Berezin et al., 2004</xref>). The <italic>sap130a</italic> amino acid sequence from zebrafish was input to ConSurf and the output was collected and plotted in R, with ggtree, ggplot2 and phytools (<xref ref-type="bibr" rid="B72">Revell and Graham Reynolds, 2012</xref>; <xref ref-type="bibr" rid="B88">Wickham, 2016</xref>; <xref ref-type="bibr" rid="B92">Yu et al., 2018</xref>; <xref ref-type="bibr" rid="B93">Yu, 2020</xref>). A multiple sequence alignment (MSA) was performed on Sap130 protein sequences from UniProt and distance calculations to plot simple phylogeny trees using R CRAN packages seqinr, msa, Biostrings, ggtree, ggplot2 (<xref ref-type="bibr" rid="B13">Charif et al., 2005</xref>; <xref ref-type="bibr" rid="B8">Bodenhofer et al., 2015</xref>; <xref ref-type="bibr" rid="B56">Lifschitz et al., 2022</xref>). For plotting the protein sequences and conserved domains reported by UniProt, the R packages ggplot and drawProteins were used (<xref ref-type="bibr" rid="B9">Brennan, 2018</xref>).</p>
</sec>
<sec id="s2-5">
<title>
<italic>In situ</italic> probe synthesis, whole mount <italic>in situ</italic> hybridization</title>
<p>RNA probe generation and whole mount <italic>in situ</italic> hybridization for <italic>nkx2.5</italic>, <italic>ltbp3</italic>, <italic>myh7</italic> and <italic>myh6</italic> was performed as previously described with DIG RNA labeling kit (Millipore Sigma cat&#x23; 11175025910) (<xref ref-type="bibr" rid="B96">Znosko et al., 2010</xref>).</p>
</sec>
<sec id="s2-6">
<title>RNAseq sample preparation and data analysis</title>
<p>Total RNA was extracted from whole embryos or isolated hearts (36hpf and 48hpf, respectively) using Trizol (Invitrogen) and was purified with the RNeasy Micro Kit (Qiagen&#x23;74004). A minimum 50 embryos or 180 hearts were pooled together for each condition. The RNA-seq used was 0.5&#x2013;1&#xa0;&#x3bc;g RNA for each condition and was sent to the Genomics Research Core at the University of Pittsburgh. The raw sequence reads were processed and mapped to the Zebrafish Reference Genome GRCz11 using CLC Genomics Workbench 20 RNAseq analysis tool. A count matrix was exported and the bioinformatic analysis was carried out in R (<xref ref-type="bibr" rid="B71">R Core Team, 2021</xref>) using the edgeR package for 36hpf whole embryo and 48hpf heart tissue data. Results for DEGs in <xref ref-type="sec" rid="s11">Supplementary Tables, S2&#x2013;S6</xref> (<xref ref-type="bibr" rid="B73">Robinson et al., 2010</xref>). To identify cardiac changes with whole embryo resolution we defined DEGs as those with an FDR &#x2264; 0.05 and log2FC &#x3e; &#xb1;0.4. After determining differentially expressed genes they were entered into DAVID (<ext-link ext-link-type="uri" xlink:href="https://david.ncifcrf.gov/summary.jsp">https://david.ncifcrf.gov/summary.jsp</ext-link>) for functional annotation clustering. Results for DAVID clustering in <xref ref-type="sec" rid="s11">Supplementary Tables, S2&#x2013;S6</xref> (<xref ref-type="bibr" rid="B76">Sherman et al., 2022</xref>).</p>
</sec>
<sec id="s2-7">
<title>Lineage tracing</title>
<p>Lineage tracing of cardiac progenitors at 24hpf was performed on <italic>Tg(nkx2.5:kaede)</italic> and <italic>Tg(nkx2.5:kaede);sap130a</italic>
<sup>
<italic>pt32a/pt32a</italic>
</sup> embryos and was described by Guner-Ataman et al. (<xref ref-type="bibr" rid="B31">Guner-Ataman et al., 2013</xref>). Using the Zeiss Imager M2 confocal microscope at 40x, the ROI (Region of Interest) was selected to photoconvert the peristaltic heart tube at 24hpf. Embryos were mounted in low melting temperature agarose droplets on 35&#xa0;mm dishes. The embryos were then freed from the agarose and raised in darkness until 48hpf, when the looped heart was imaged at 40x.</p>
</sec>
<sec id="s2-8">
<title>Cardiac functional analysis</title>
<p>To measure cardiac function in embryonic zebrafish, we used a custom-built light sheet microscope which followed a design based on the openSPIM platform (<xref ref-type="bibr" rid="B68">Pitrone et al., 2013</xref>; <xref ref-type="bibr" rid="B28">Girstmair et al., 2016</xref>). This &#x2018;T&#x2019; design illuminates the sample bilaterally and uses a four-channel laser launch for maximum versatility. <italic>Tg(myl7:EGFP)</italic> and <italic>Tg(myl7:EGFP);sap130a</italic>
<sup>
<italic>m/m</italic>
</sup> embryos at 48hpf embryos were placed into E3 and Tricaine (307&#xa0;nmol concentration) to anesthetize them before mounting for imaging. Low melting point agarose was heated and cooled to 42&#xb0;C. 100&#xa0;&#xb5;L agarose placed onto a dish and after 45&#xa0;s of cooling, 48hpf embryo was added to the agarose and drawn into a custom cut 1&#xa0;mL straight-barreled syringe. The agarose is allowed to solidify, and the syringe is placed into a sample manipulator capable of 3D movement &#x2b; rotation (Picard Technologies, Inc.). The agarose-embedded embryos were extruded from the syringe and positioned in a lateral view, with anterior to the left and posterior to the right, before recording 100 frames at 50&#x2013;75 frames per second using a Prime 95B sCMOS camera (Photometrics, Inc.). Fiji ImageJ software was used to identify end-diastole and end-systole frames to calculate ventricle area, length (distance between ventricular apex and out-flow tract opening), and diameter for each embryo (distance between the walls of the chamber, taken from the middle of length measurement). These data were used to estimate chamber volumes and calculate end-diastole and systole volumes, ejection fraction (%), fractional shortening (&#xb5;m), Total stroke volume, cardiac output, and heart rate as an average of all cycles captured for each fish. The volumes calculated are under the assumption of a prolate sphere shape (pi/6). The equations used are as follows (<xref ref-type="bibr" rid="B91">Yalcin et al., 2017</xref>);<disp-formula id="equ1">
<mml:math id="m1">
<mml:mrow>
<mml:mi>E</mml:mi>
<mml:mi>j</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>c</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>n</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>F</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>c</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>n</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mrow>
<mml:mfenced open="(" close=")" separators="|">
<mml:mrow>
<mml:mo>%</mml:mo>
</mml:mrow>
</mml:mfenced>
</mml:mrow>
<mml:mo>&#x3d;</mml:mo>
<mml:mfrac>
<mml:mrow>
<mml:mi>S</mml:mi>
<mml:mi>V</mml:mi>
</mml:mrow>
<mml:mrow>
<mml:mi>E</mml:mi>
<mml:mi>D</mml:mi>
<mml:mi>V</mml:mi>
</mml:mrow>
</mml:mfrac>
<mml:mo>&#x2a;</mml:mo>
<mml:mn>100</mml:mn>
</mml:mrow>
</mml:math>
</disp-formula>
<disp-formula id="equ2">
<mml:math id="m2">
<mml:mrow>
<mml:mi>E</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>d</mml:mi>
<mml:mo>_</mml:mo>
<mml:mi>D</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mo>&#x26;</mml:mo>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>E</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>d</mml:mi>
<mml:mo>_</mml:mo>
<mml:mi>S</mml:mi>
<mml:mi>y</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>V</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>u</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>s</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mrow>
<mml:mfenced open="(" close=")" separators="|">
<mml:mrow>
<mml:mi>E</mml:mi>
<mml:mi>D</mml:mi>
<mml:mi>V</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mo>&#x26;</mml:mo>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>E</mml:mi>
<mml:mi>S</mml:mi>
<mml:mi>V</mml:mi>
</mml:mrow>
</mml:mfenced>
</mml:mrow>
<mml:mo>&#x3d;</mml:mo>
<mml:mfrac>
<mml:mrow>
<mml:mi>&#x3c0;</mml:mi>
</mml:mrow>
<mml:mrow>
<mml:mn>6</mml:mn>
</mml:mrow>
</mml:mfrac>
<mml:mo>&#x2a;</mml:mo>
<mml:mi>L</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>g</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>h</mml:mi>
<mml:mo>&#x2a;</mml:mo>
<mml:msup>
<mml:mrow>
<mml:mi>D</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>r</mml:mi>
</mml:mrow>
<mml:mn>2</mml:mn>
</mml:msup>
</mml:mrow>
</mml:math>
</disp-formula>
<disp-formula id="equ3">
<mml:math id="m3">
<mml:mrow>
<mml:mi>S</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>k</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>V</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>u</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mrow>
<mml:mfenced open="(" close=")" separators="|">
<mml:mrow>
<mml:mi>S</mml:mi>
<mml:mi>V</mml:mi>
</mml:mrow>
</mml:mfenced>
</mml:mrow>
<mml:mo>&#x3d;</mml:mo>
<mml:mi>E</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>d</mml:mi>
<mml:mo>_</mml:mo>
<mml:mi>D</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>V</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>u</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
<mml:mo>&#x2212;</mml:mo>
<mml:mi>E</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>d</mml:mi>
<mml:mo>_</mml:mo>
<mml:mi>S</mml:mi>
<mml:mi>y</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>V</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>u</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
</mml:mrow>
</mml:math>
</disp-formula>
<disp-formula id="equ4">
<mml:math id="m4">
<mml:mrow>
<mml:mi>F</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>c</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>l</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>S</mml:mi>
<mml:mi>h</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>g</mml:mi>
<mml:mo>&#x3d;</mml:mo>
<mml:mfrac>
<mml:mrow>
<mml:mi>D</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>d</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>r</mml:mi>
<mml:mo>&#x2212;</mml:mo>
<mml:mi>S</mml:mi>
<mml:mi>y</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>d</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>r</mml:mi>
</mml:mrow>
<mml:mrow>
<mml:mi>D</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi mathvariant="normal">d</mml:mi>
<mml:mi mathvariant="normal">i</mml:mi>
<mml:mi mathvariant="normal">a</mml:mi>
<mml:mi mathvariant="normal">m</mml:mi>
<mml:mi mathvariant="normal">e</mml:mi>
<mml:mi mathvariant="normal">t</mml:mi>
<mml:mi mathvariant="normal">e</mml:mi>
<mml:mi mathvariant="normal">r</mml:mi>
</mml:mrow>
</mml:mfrac>
</mml:mrow>
</mml:math>
</disp-formula>
<disp-formula id="equ5">
<mml:math id="m5">
<mml:mrow>
<mml:mi>F</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>c</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>l</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>A</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>a</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>C</mml:mi>
<mml:mi>h</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>g</mml:mi>
<mml:mi>e</mml:mi>
<mml:mo>&#x3d;</mml:mo>
<mml:mfrac>
<mml:mrow>
<mml:mi>E</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>d</mml:mi>
<mml:mo>_</mml:mo>
<mml:mi>D</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>A</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>a</mml:mi>
<mml:mo>&#x2212;</mml:mo>
<mml:mi>E</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>d</mml:mi>
<mml:mo>_</mml:mo>
<mml:mi>S</mml:mi>
<mml:mi>y</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>A</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>a</mml:mi>
</mml:mrow>
<mml:mrow>
<mml:mi>E</mml:mi>
<mml:mi>n</mml:mi>
<mml:mi>d</mml:mi>
<mml:mo>_</mml:mo>
<mml:mi>D</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>A</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>a</mml:mi>
</mml:mrow>
</mml:mfrac>
<mml:mo>&#x2a;</mml:mo>
<mml:mn>100</mml:mn>
</mml:mrow>
</mml:math>
</disp-formula>
<disp-formula id="equ6">
<mml:math id="m6">
<mml:mrow>
<mml:mi>H</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>r</mml:mi>
<mml:mi>t</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>R</mml:mi>
<mml:mi>a</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>e</mml:mi>
<mml:mo>&#x3d;</mml:mo>
<mml:mfrac>
<mml:mrow>
<mml:mo>&#x23;</mml:mo>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>o</mml:mi>
<mml:mi>f</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>C</mml:mi>
<mml:mi>y</mml:mi>
<mml:mi>c</mml:mi>
<mml:mi>l</mml:mi>
<mml:mi>e</mml:mi>
<mml:mi>s</mml:mi>
</mml:mrow>
<mml:mrow>
<mml:mi>A</mml:mi>
<mml:mi>q</mml:mi>
<mml:mi>u</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>s</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>t</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>o</mml:mi>
<mml:mi>n</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mi>t</mml:mi>
<mml:mi>i</mml:mi>
<mml:mi>m</mml:mi>
<mml:mi>e</mml:mi>
<mml:mtext>&#x2009;</mml:mtext>
<mml:mrow>
<mml:mfenced open="(" close=")" separators="|">
<mml:mrow>
<mml:mi>s</mml:mi>
</mml:mrow>
</mml:mfenced>
</mml:mrow>
</mml:mrow>
</mml:mfrac>
</mml:mrow>
</mml:math>
</disp-formula>
</p>
<p>These were implemented using R scripting and RStudio to automate the calculations, and then data were plotted using Graphpad PRISM 9.3. Each data point represents an average of 3 or more contraction cycles per fish (<xref ref-type="bibr" rid="B91">Yalcin et al., 2017</xref>).</p>
</sec>
<sec id="s2-9">
<title>Adult heart measurements</title>
<p>At 48hpf <italic>MZsap130a</italic> mutant embryos were scored for ventricle size and raised in separate tanks. <italic>MZsap130a</italic> mutants and aged matched <italic>AB&#x2a;</italic> controls were measured for length and weight before hearts were extracted for DIC imaging at 4-6mpf. Fiji-ImageJ was used to measure the ventricle surface area and bulbus arteriosus surface area. These data were plotted using Graphpad Prism 9.3.</p>
</sec>
<sec id="s2-10">
<title>Statistics</title>
<p>For analysis of RNA-seq data we used the edgeR package, utilizing a quasi-likelihood negative binomial generalized log-linear model to our count data comparing AB&#x2a; control to <italic>MZsap130a</italic> or <italic>MZsap130b</italic> mutant embryos at 36hpf. For heart tissue RNA-seq, edgeR&#x2019;s likelihood ratio test was used to interpret up or downregulation of genes. For all other statistical analysis, significance was calculated using two-tailed, unpaired Student&#x2019;s t-test, one-way ANOVA or Fisher&#x2019;s exact text using GraphPad Prism version 9.3.</p>
</sec>
</sec>
<sec sec-type="results" id="s3">
<title>Results</title>
<sec id="s3-1">
<title>
<italic>sap130b</italic> is not required for heart development</title>
<p>Zebrafish were part of the teleost-specific genome duplication event 350 million years ago (<xref ref-type="bibr" rid="B3">Alsop and Vijayan, 2009</xref>), resulting in two <italic>sap130</italic> genes, <italic>sap130a</italic> and <italic>sap130b</italic>. Defining the SAP130 protein domains based on homology with other model organisms will provide insight into the potential conserved functional domains. In mammals, both SIN3A and HDAC1 proteins were shown to interact with SAP130 at the C-terminus between amino acids 836&#x2013;1,047, suggesting that SAP130 may act as a stabilizing scaffold between these proteins (<xref ref-type="bibr" rid="B25">Fleischer et al., 2003</xref>). Determining protein sequence similarities can predict functional structures across species and offer insight into the potential for functional redundancy between Sap130a and Sap130b. ConSurf was used for a multispecies comparison of 145 unique SAP130 protein sequences to determine their similarity and conserved domains (<xref ref-type="bibr" rid="B6">Berezin et al., 2004</xref>). In general, Sap130a and Sap130b are dissimilar, but they both contained conserved N- and C-terminus domains represented by repetitive predicted structural and functional residues (<xref ref-type="sec" rid="s11">Supplementary Figure S1</xref>). Comparing SAP130 proteins to a small group of common species Sap130a and Sap130b are most like one another, suggesting they could serve similar functions (<xref ref-type="fig" rid="F1">Figure 1A</xref>). Narrowing the comparison to a smaller set of protein sequences among other teleost, Sap130a and Sap130b are distinct suggesting in teleost these genes could have evolved distinct functions (<xref ref-type="fig" rid="F1">Figure 1B</xref>). However, given that the C- terminal domains are most conserved, Sap130a and Sap130b can potentially compensate for one another in zebrafish (<xref ref-type="fig" rid="F1">Figure 1C</xref>). <italic>MZsap130a</italic> mutants develop SVs in 36% of the population by 72hpf (<xref ref-type="bibr" rid="B57">Liu et al., 2017</xref>). The incomplete penetrance of the SV phenotype was hypothesized to be the result of <italic>sap130b</italic> compensating for the loss of <italic>sap130a</italic>. To address this, we generated a mutation in <italic>sap130b</italic> using CRISPR/Cas9 technology. This produced an allele (7bp del, 1bp sub (G&#x3e;C))<italic>sap130b</italic>
<sup>
<italic>pt35b/pt35b</italic>
</sup> that introduced a premature stop codon in exon 6 of <italic>sap130b</italic> disrupting the N-terminus and eliminating the C-terminal region (<xref ref-type="fig" rid="F1">Figure 1C</xref>, <xref ref-type="sec" rid="s11">Supplementary Tables S1</xref>). Using the <italic>Tg(myl7:EGFP)</italic> line, which labels the heart with green fluorescent protein, we found that 48% of the <italic>MZsap130a;Tg(myl7:EGFP)</italic> mutant embryos had the SV heart phenotype at 48hpf (<xref ref-type="fig" rid="F1">Figure 1D</xref>). In contrast, only 17% of the <italic>MZsap130b;Tg(myl7:EGFP)</italic> mutant embryos had SVs by 48hpf (<xref ref-type="fig" rid="F1">Figure 1E</xref>). We generated double mutants to further explore if <italic>sap130a</italic> and <italic>sap130b</italic> have any redundant functions (<xref ref-type="fig" rid="F2">Figures 2A, B</xref>). The offspring produced the expected number of double mutants (7/120 (5.8%)) from the expected (1/16 (6.25%)) from a double heterozygous in-cross. However, the adult double <italic>sap130a/b</italic> mutants are much smaller than their double heterozygous siblings and failed to produce offspring when bred (<xref ref-type="fig" rid="F2">Figure 2C</xref>). <italic>MZsap130a;sap130b</italic>
<sup>
<italic>pt35b/&#x2b;</italic>
</sup> mutant in-crosses, resulted in 39% of the embryos with SVs at 48hpf, which is in the same range as <italic>MZsap130a</italic> mutants indicating the zygotic loss of <italic>sap130b</italic> did not contribute to increased cardiac defects (<xref ref-type="fig" rid="F2">Figure 2D</xref>). These observations suggest <italic>sap130b</italic> is not required for zebrafish cardiogenesis.</p>
<fig id="F1" position="float">
<label>FIGURE 1</label>
<caption>
<p>
<italic>sap130a</italic> and <italic>sap130b</italic> have non-overlapping functions in the zebrafish heart <bold>(A, B)</bold> A simple distance matrix phylogeny tree of Sap130a and Sap130b in broad or teleost specific contexts <bold>(C)</bold> Schematic of Sap130a and Sap130b protein sequences from the UniProt database highlighting the conserved regions and predicted mutant proteins. Unorganized sequence in pink, C-terminal conserved domain in blue, which contains the binding domain for SIN3A and HDAC1 <bold>(D, E)</bold> Representative images of <italic>Tg</italic>(<italic>myl7:EGFP</italic>), <italic>MZsap130a;Tg(myl7:EGFP)</italic> and <italic>MZsap130b;Tg(myl7:EGFP)</italic> mutant hearts at 48hpf. V and A are ventricle and atria, respectively. Scale bar 100&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g001.tif"/>
</fig>
<fig id="F2" position="float">
<label>FIGURE 2</label>
<caption>
<p>
<italic>MZsap130a;sap130b</italic>
<sup>
<italic>pt35b/pt35b</italic>
</sup> mutants are not healthy <bold>(A)</bold> <italic>sap130a</italic>;<italic>sap130b</italic> double heterozygous adults, male (top) and female (bottom). <bold>(B)</bold> <italic>sap130a</italic>;<italic>sap130b</italic> double homozygous adults, male (top) and female (bottom). <bold>(C)</bold> Graph quantifying weight to length ratio for adults from a <italic>sap130a</italic>;<italic>sap130b</italic> double heterozygous in-cross, pvals are for one-way ANOVA, error bars are standard error mean (SEM). Red points represent females and males in black. <bold>(D)</bold> Graph quantifying the heart phenotype proportions for <italic>Tg</italic>(<italic>myl7:EGFP</italic>), <italic>MZsap130a;Tg(myl7:EGFP)</italic>, <italic>MZsap130b;Tg</italic>(<italic>myl7:EGFP</italic>), and <italic>MZsap130a;sap130b</italic>
<sup>
<italic>pt35b/&#x2b;</italic>
</sup>
<italic>;Tg</italic>(<italic>myl7:EGFP</italic>)<bold>,</bold> pvals are for fisher&#x2019;s exact test. Scale bar 5&#xa0;mm.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g002.tif"/>
</fig>
<p>
<italic>Sap130a</italic> AUG start codon antisense-morpholino (MO) studies suggested the SVs arise from decreased ventricular CMs (<xref ref-type="bibr" rid="B57">Liu et al., 2017</xref>), but where or when CMs are lost was not explored. To determine if the SVs are due to decreased cardiac progenitors, we performed Whole Mount <italic>In Situ</italic> Hybridization (WISH) at 10 somite stage with <italic>nkx2.5</italic>, an early cardiac progenitor marker. We discovered no differences between <italic>MZsap130a</italic> and controls (<xref ref-type="fig" rid="F3">Figure 3A</xref>). This suggests that the early cardiac progenitors were present in the <italic>MZsap130a</italic> embryos. To profile a later stage of the First Heart Field (FHF) and the chambers of the heart we performed WISH at 24hpf with myosin heavy chain 7 (<italic>myh7</italic>, ventricle) and myosin heavy chain 6 (<italic>myh6</italic>, atria). No difference between WT and mutant embryos were observed, suggesting the FHF is intact (<xref ref-type="fig" rid="F3">Figure 3B</xref>). At 36hpf and 48hpf the atrial chamber showed no change, but the ventricle was smaller (<xref ref-type="fig" rid="F3">Figure 3C</xref>, <xref ref-type="sec" rid="s11">Supplementary Figure S2</xref>). This phenotype was observed again when imaging the <italic>MZsap130a;Tg(myl7:EGFP)</italic> at 36hpf (<xref ref-type="fig" rid="F4">Figure 4A</xref>). Many studies have detailed the second heart field accretion between 24 and 48hpf in zebrafish (<xref ref-type="bibr" rid="B30">Grimes et al., 2008</xref>; <xref ref-type="bibr" rid="B20">de Pater et al., 2009</xref>; <xref ref-type="bibr" rid="B32">Hami et al., 2011</xref>; <xref ref-type="bibr" rid="B52">Lazic and Scott, 2011</xref>). These SHF cells trail behind the heart tube and add to the ventricle continuously. There is speculation as to how many SHF cells are ventricular CMs, between 30%&#x2013;40% of the total ventricular CMs by 48hpf has been proposed (<xref ref-type="bibr" rid="B24">Felker et al., 2018</xref>). The SV heart phenotype arising at 36hpf and the lack of changes seen in FHF markers suggest the SHF might be an influenced cell population where CMs are lost in <italic>MZsap130a</italic> mutants.</p>
<fig id="F3" position="float">
<label>FIGURE 3</label>
<caption>
<p>Cardiac gene expression in <italic>MZsap130a</italic> and <italic>MZsap130b</italic> <bold>(A)</bold> WISH of <italic>nkx2.5</italic>&#xa0;at 10 somite stage for <italic>AB&#x2a;</italic>, <italic>MZsap130a</italic> and <italic>MZsap130b</italic>. <bold>(B)</bold> WISH of <italic>myh6</italic> and <italic>myh7</italic> at 24hpf for <italic>AB&#x2a;</italic>, <italic>MZsap130a</italic> and <italic>MZsap130b</italic>. <bold>(C)</bold> WISH of <italic>myh7</italic> at 36hpf in <italic>AB&#x2a;</italic> and <italic>MZsap130a</italic>. Scale bar 100&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g003.tif"/>
</fig>
<fig id="F4" position="float">
<label>FIGURE 4</label>
<caption>
<p>RNAseq reveals cardiac contraction and conduction is altered in <italic>MZsap130a</italic> <bold>(A)</bold> Representative images of <italic>Tg</italic>(<italic>myl7:EGFP</italic>) and <italic>MZsap130a;Tg(myl7:EGFP)</italic> embryos collected for whole embryo RNAseq at 36hpf. <bold>(B)</bold> Volcano plot of 36hpf whole embryo RNAseq data. <bold>(C)</bold> Heatmap of sarcomere and conduction genes (<xref ref-type="sec" rid="s11">Supplementary Figure S6</xref>) at 36hpf from whole embryos. <bold>(D)</bold> DAVID functional annotation cluster 8 from downregulated genes in <italic>MZsap130a</italic>, showing DAVID calculated <italic>p</italic>-values. <bold>(E)</bold> Heatmap of 48hpf heart tissue RNAseq data for the same genes found in panel C. V and A are ventricle and atria, respectively. Scale bar 100&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g004.tif"/>
</fig>
</sec>
<sec id="s3-2">
<title>RNAseq reveals <italic>sap130a</italic> is involved in regulating cardiac sarcomere and conduction genes</title>
<p>Variants in genes encoding sarcomere proteins have been linked to CHDs (<xref ref-type="bibr" rid="B46">Jones et al., 1996</xref>; <xref ref-type="bibr" rid="B62">Morano et al., 2000</xref>; <xref ref-type="bibr" rid="B16">Ching et al., 2005</xref>; <xref ref-type="bibr" rid="B95">Zhu et al., 2006</xref>; <xref ref-type="bibr" rid="B60">Monserrat et al., 2007</xref>; <xref ref-type="bibr" rid="B29">Granados-Riveron et al., 2010</xref>; <xref ref-type="bibr" rid="B69">Postma et al., 2011</xref>; <xref ref-type="bibr" rid="B23">Fahed et al., 2013</xref>; <xref ref-type="bibr" rid="B83">van Engelen et al., 2013</xref>). The Sin3A complex has been shown to regulate sarcomere specific genes like titins, troponins, and actins important for cardiac contraction (<xref ref-type="bibr" rid="B84">van Oevelen et al., 2010</xref>). Since SAP130 has been shown to be part of the SIN3A complex, we reasoned that the phenotype may be caused by altered regulation of cardiac gene expression during development. A whole embryo RNA sequencing (RNAseq) experiment, separating the SV and &#x201c;normal&#x201d; (NV) siblings in the <italic>MZsap130a</italic> mutants, was performed at 36hpf. We first performed our analysis looking for differences in the wildtype, compared to NV and SV separately finding 2,826 differentially expressed genes (DEGs) in common, with 812 unique DEGs for NV and 1979 for SV. Functional annotation of these gene groups revealed that NV and SV embryos are similar when compared to the wildtype transcriptome (<xref ref-type="sec" rid="s11">Supplementary Table S2</xref>). Comparing the controls to all <italic>MZsap130a</italic> samples (both NV and SV), we observed 5,002 DEGs that included many cardiac specific transcripts. Among the DEGs we found sarcomere and cardiac conduction genes were dysregulated, suggesting CM biology has changed in the <italic>MZsap130a</italic> embryos (<xref ref-type="fig" rid="F4">Figures 4B, C</xref>, <xref ref-type="sec" rid="s11">Supplementary Tables S3, S4</xref>). To identify potential pathways involved in heart function and development, we used the Database for Annotation, Visualization, and Integrated Discovery (DAVID) functional annotation of downregulated genes. This showed enrichment for cardiac contraction and adrenergic signaling in CMs, further suggesting a role for <italic>sap130a</italic> in CM function (<xref ref-type="fig" rid="F4">Figure 4D</xref>, <xref ref-type="sec" rid="s11">Supplementary Table S6</xref>). To confirm cardiac specific changes in these same transcripts, <italic>MZsap130a</italic> mutant hearts and controls were harvested at 48hpf and the transcriptome was profiled, showing similar cardiac gene expression changes (<xref ref-type="fig" rid="F4">Figure 4E</xref>, <xref ref-type="sec" rid="s11">Supplementary Table S5</xref>). <italic>MZsap130b</italic> whole embryo transcriptome was also profiled at 36hpf and less gene expression changes (617 DEGs) were noted (<xref ref-type="sec" rid="s11">Supplementary Figure S3</xref>, <xref ref-type="sec" rid="s11">Supplementary Table S6</xref>). Moreover, the sarcomere gene expression changes seen in <italic>MZsap130a</italic> was not detected in the <italic>MZsap130b</italic> transcriptome. DAVID functional annotation of the 278 DEGs common between <italic>MZsap130a</italic> and <italic>MZsap130b</italic> mutants, belonged to heme binding and biosynthesis, oxygen binding, and iron binding KEGG pathways, suggesting involvement in hematopoiesis (<xref ref-type="sec" rid="s11">Supplementary Table S6</xref>). The expression profile for these hematopoietic related genes was opposite in <italic>MZsap130a</italic> and <italic>MZsap130b</italic>, suggesting distinct functions during hematopoiesis (<xref ref-type="sec" rid="s11">Supplementary Figure S3</xref>). These data suggest that <italic>sap130a</italic> and <italic>sap130b</italic> could be involved in hematopoiesis that correlates with <italic>sin3aa</italic>/<italic>ab</italic> gene knockdown studies showing strong hematopoietic defects (<xref ref-type="bibr" rid="B39">Huang et al., 2013</xref>).</p>
<p>Whole embryo and heart tissue <italic>MZsap130a</italic> RNA-seq data revealed sarcomere genes such as actins and myosins were dysregulated, indicating that sarcomere dysfunction could be part for the <italic>MZsap130a</italic> mutant phenotype. These data also showed downregulation of CM conduction genes such as <italic>cxcr4b</italic> and <italic>gja3</italic>, resulting in changes in cardiogenesis (<xref ref-type="bibr" rid="B74">Severs et al., 2004</xref>; <xref ref-type="bibr" rid="B79">Stankunas et al., 2008</xref>; <xref ref-type="bibr" rid="B15">Chi et al., 2010</xref>; <xref ref-type="bibr" rid="B42">Itou et al., 2012</xref>; <xref ref-type="bibr" rid="B63">Mortensen et al., 2017</xref>; <xref ref-type="bibr" rid="B44">Jiang et al., 2019</xref>). <italic>dococ</italic>
<sup>
<italic>s226</italic>
</sup> (<italic>gja3</italic>) mutants report having changes in cardiac conduction that lead to CM morphological changes in the ventricle (<xref ref-type="bibr" rid="B15">Chi et al., 2010</xref>). Rat studies have shown <italic>Cxcr4</italic> involvement in cardiac conduction (<xref ref-type="bibr" rid="B70">Pyo et al., 2006</xref>). While <italic>MZcxcr4b</italic> mutants are reported to have abnormal organ morphogenesis, including heart looping defects (<xref ref-type="bibr" rid="B44">Jiang et al., 2019</xref>). Changes were found in calcium channel (<italic>cacna1sb</italic>, <italic>cacng7a</italic>, <italic>cacnb1</italic>, <italic>cacna1bb</italic>) and sodium channel (<italic>scn4aa</italic>/<italic>ab</italic>, <italic>scn2b</italic>) genes, known to be important to CM biology (<xref ref-type="bibr" rid="B36">Haverinen et al., 2018</xref>; <xref ref-type="bibr" rid="B67">Papa et al., 2022</xref>; <xref ref-type="bibr" rid="B75">Shah et al., 2022</xref>). Furthermore, transcriptome analysis revealed that <italic>MZsap130a</italic> mutants showed dysregulation of a wide range of genes critical for cardiac maturation and function. These include genes associated with fatty acid metabolism (<italic>ppt2</italic>), glycogen metabolism (<italic>ugp2a</italic>, <italic>phka2</italic>), and mitochondria (<italic>slc25a44a</italic>, <italic>slc25a42</italic>, <italic>mtrf1</italic>, <italic>mrpl58</italic>) found downregulated in <italic>MZsap130a</italic> mutants in whole embryos at 36hpf and specifically in the heart at 48hpf (<xref ref-type="sec" rid="s11">Supplementary Figure S3</xref>). Deficits in mitochondrial function have been shown in HLHS patients and other HLHS models including <italic>Sap130</italic> mouse mutants and <italic>rbfox</italic> mutant zebrafish (<xref ref-type="bibr" rid="B57">Liu et al., 2017</xref>; <xref ref-type="bibr" rid="B40">Huang et al., 2022</xref>). Collectively, <italic>MZsap130a</italic> mutants show changes in sarcomere, conduction and metabolism associated genes, all integral parts of CM maturation and function.</p>
</sec>
<sec id="s3-3">
<title>Sap130a regulates cardiac function</title>
<p>Global loss of <italic>sap130a</italic> showed downregulation of sarcomere genes such as <italic>actc1</italic>, <italic>ttn.1</italic>, and <italic>ttn.2</italic> (<xref ref-type="fig" rid="F4">Figures 4B&#x2013;E</xref>
<bold>)</bold>. This suggested that cardiac function could be diminished in <italic>MZsap130a</italic> mutants. The DAVID functional annotation tool revealed enrichment for cardiac muscle contraction genes that were decreased in the <italic>MZsap130a</italic> mutant embryos (<xref ref-type="fig" rid="F4">Figure 4D</xref>). To determine ventricle chamber function in mutants, confocal light sheet microscopy was used to record live cardiac contractions at 48hpf. These recordings provided us with multiple frames of diastole and systole for chamber volume estimation (<xref ref-type="fig" rid="F5">Figures 5A&#x2013;C</xref>, and <xref ref-type="sec" rid="s11">Supplementary Movie S1&#x2013;S5</xref>). Volume estimations were used to calculate the cardiac parameters Total Stroke Volume (TSV), and Cardiac Output (CO) (<xref ref-type="bibr" rid="B91">Yalcin et al., 2017</xref>). The light sheet data revealed that all <italic>MZsap130a</italic> mutants had deficits in CO, TSV, fractional shortening, and ejection fraction (<xref ref-type="fig" rid="F5">Figures 5D, E</xref>, and <xref ref-type="sec" rid="s11">Supplementary Figure S5</xref>). The <italic>MZsap130b</italic> mutant hearts revealed no significant difference from WT function, both in TSV and CO, but showed an increase in End-systolic volume which could explain the increase in CO through increased contraction force (<xref ref-type="fig" rid="F5">Figures 5D, E</xref> and <xref ref-type="sec" rid="s11">Supplementary Figure S5</xref>). The heart tissue RNA-seq identified cardiac contraction genes <italic>myh7</italic>, <italic>actc1</italic>, <italic>ttn.1</italic>, <italic>ttn.2</italic>, <italic>scn4ab</italic>, and <italic>cacna1sb</italic> were dysregulated in <italic>MZsap130a</italic> mutants, supporting the contraction deficits measured at 48hpf (<xref ref-type="sec" rid="s11">Supplementary Figure S6</xref>). These data show that <italic>sap130a</italic> has a role in zebrafish cardiac sarcomere regulation.</p>
<fig id="F5" position="float">
<label>FIGURE 5</label>
<caption>
<p>
<italic>MZsap130a</italic> show cardiac functional deficits <bold>(A&#x2013;C)</bold> Shows systole and diastole frames from recordings of live ventricular contractions in <italic>Tg</italic>(<italic>myl7:EGFP</italic>), <italic>MZsap130a;Tg(myl7:EGFP)</italic> and <italic>MZsap130b;Tg</italic>(<italic>myl7:EGFP</italic>) at 48hpf. <bold>(D, E)</bold> Quantified cardiac parameters total stroke volume (TSV) and cardiac output (CO), pvals from one-way ANOVA, error bars are SEM. Each point represents individual ventricle and color coded for 3&#x2b; experiments. For <italic>Tg(myl7:EGFP)</italic>, <italic>n</italic> &#x3d; 115; <italic>MZsap130a</italic> NV, <italic>n</italic> &#x3d; 36; <italic>MZsap130a</italic> SV, <italic>n</italic> &#x3d; 30; <italic>MZsap130b</italic> NV, <italic>n</italic> &#x3d; 57; <italic>MZsap130b</italic> SV, <italic>n</italic> &#x3d; 13. Scale bar 20&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g005.tif"/>
</fig>
</sec>
<sec id="s3-4">
<title>
<italic>MZsap130a</italic> mutants have longer outflow tract</title>
<p>The earliest observation of smaller ventricles in <italic>MZsap130a</italic> mutants was at 36hpf, a stage when SHF cells are migrating into the ventricle. Extensive studies have reported the contribution of SHF cells to the ventricle during this time (<xref ref-type="bibr" rid="B30">Grimes et al., 2008</xref>; <xref ref-type="bibr" rid="B20">de Pater et al., 2009</xref>; <xref ref-type="bibr" rid="B52">Lazic and Scott, 2011</xref>; <xref ref-type="bibr" rid="B50">Knight and Yelon, 2016</xref>; <xref ref-type="bibr" rid="B24">Felker et al., 2018</xref>; <xref ref-type="bibr" rid="B78">Song et al., 2019</xref>). RNA-seq data showed that SHF progenitor markers <italic>ltbp3</italic>, <italic>mef2cb</italic> and <italic>isl1, isl2a/b</italic> were decreased (<xref ref-type="sec" rid="s11">Supplementary Figure S7</xref>). These genes are known to label SHF progenitors at the arterial and venous poles. WISH at 30hpf revealed a decrease in <italic>ltbp3</italic> expression in <italic>MZsap130a</italic> mutants (<xref ref-type="sec" rid="s11">Supplementary Figure S8</xref>). Together these data suggest that the SHF in the <italic>MZsap130a</italic> mutants was affected such that insufficient CMs contribute to the ventricle by 48hpf. To determine if this occurs, we performed lineage tracing experiments using <italic>Tg(nkx2.5:kaede)</italic> embryos (<xref ref-type="bibr" rid="B31">Guner-Ataman et al., 2013</xref>). In this transgenic line, the FHF cells can be permanently labeled at 24hpf, photo-converting only the heart tube. Next, we imaged at 48hpf to determine the addition of green cells to the ventricle (<xref ref-type="fig" rid="F6">Figure 6A</xref>, and <xref ref-type="sec" rid="s11">Supplementary Figure S9</xref>. Lineage tracing the SHF with <italic>MZsap130a;Tg(nkx2.5:kaede)</italic> embryos revealed that the SVs acquire less SHF (green area) compared to the wildtype and <italic>MZsap130a</italic> mutant siblings that develop normal ventricles (<xref ref-type="fig" rid="F6">Figures 6B&#x2013;D</xref>). Moreover, the OFTs in the <italic>MZsap130a</italic> mutants were longer at 48hpf in some embryos with SVs (<xref ref-type="sec" rid="s11">Supplementary Figure S10</xref>). The longer OFTs were much more pronounced at 72hpf, and every SV heart had a longer OFT (<xref ref-type="fig" rid="F7">Figures 7A, B</xref>, and <xref ref-type="sec" rid="s11">Supplementary Figure S10</xref>). To count the CMs in the ventricle and OFT of the <italic>MZsap130a</italic> mutants, we injected <italic>Tg(myl7:memGFP)</italic> and <italic>MZsap130a;Tg(myl7:memGFP)</italic> embryos with <italic>H2b:mCherry</italic> mRNA. We detected an increase in OFT cells that was concomitant with a decrease in ventricular CMs (<xref ref-type="fig" rid="F7">Figures 7C, D</xref> and <xref ref-type="sec" rid="s11">Supplementary Figure S11</xref>). This suggested that the lost ventricular CMs contributed to OFT cells instead and was further evidenced at adult stages. <italic>MZsap130a;Tg(myl7:EGFP)</italic> embryos were scored at 48hpf for ventricle size and reared separately into adulthood. Images of heart revealed a larger bulbus arteriosus (BA) area, the adult structure derived from the OFT, and decreased ventricular area (<xref ref-type="fig" rid="F8">Figure 8</xref>). The observations in the <italic>MZsap130a</italic> adults from small ventricle embryos correlates with the notion that <italic>sap130a</italic> is involved in SHF cell fate decisions between ventricular CMs and OFT cells.</p>
<fig id="F6" position="float">
<label>FIGURE 6</label>
<caption>
<p>Lineage tracing reveal changes to SHF in <italic>MZsap130a</italic> at 48hpf <bold>(A)</bold> Diagram showing how the FHF heart tube at 24hpf was photoconverted to red, leaving SHF progenitors unlabeled in green and imaging at 48hpf. <bold>(B, C)</bold> Confocal imagines of <italic>Tg(nkx2.5:kedge)</italic> and <italic>MZsap130a;Tg(nkx2.5:kedge)</italic> at 48hpf with the heart tube being photoconverted at 24hpf, the red outlined region represents area measurements collected. <bold>(D)</bold> Quantified SHF (green area) accreted by 48hpf, pval is from a one ANOVA, error bars are SEM. Each point represents a single embryo, <italic>Tg(nkx2.5:kedge)</italic>, <italic>n</italic> &#x3d; 15; <italic>MZsap130a;Tg(nkx2.5:kedge)</italic> NV, <italic>n</italic> &#x3d; 14; <italic>MZsap130a;Tg(nkx2.5:kedge)</italic> SV, <italic>n</italic> &#x3d; 10. V and A are ventricle and atria, respectively. Scale bar 100&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g006.tif"/>
</fig>
<fig id="F7" position="float">
<label>FIGURE 7</label>
<caption>
<p>
<italic>Tg(myl7:memGFP)</italic> reveals longer OFT by 72hpf in <italic>MZsap130a</italic> <bold>(A)</bold> Quantified OFT lengths at 72hpf, pval is from a one ANOVA, SEM error bars. Each point represents a single embryo, <italic>Tg(myl7:memGFP)</italic>, <italic>n</italic> &#x3d; 18; <italic>MZsap130a;Tg(myl7:memGFP)</italic> NV, <italic>n</italic> &#x3d; 20; <italic>MZsap130a;Tg(myl7:memGFP)</italic> SV, <italic>n</italic> &#x3d; 9 <bold>(B)</bold> Representative images of <italic>Tg(myl7:memGFP)</italic> and <italic>MZsap130a;Tg(myl7:memGFP)</italic>, white lines demarcate OFT length, pvals from one way ANOVA, SEM error bars. <bold>(C)</bold> <italic>Tg(myl7:memGFP)&#x2b;;H2b:mCherry &#x2b;</italic> ventricular CM counts for three WT or <italic>MZsap130a</italic> SV heart, pval is from a t-test, SEM error bars <bold>(D)</bold> <italic>Tg(myl7:memGFP)&#x2b;;H2b:mCherry &#x2b;</italic> out-flow tract cell counts for three WT or <italic>MZsap130a</italic> SV heart, pval is from a t-test, SEM error bars. V and A are ventricle and atria, respectively. Scale bar 100&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g007.tif"/>
</fig>
<fig id="F8" position="float">
<label>FIGURE 8</label>
<caption>
<p>Adult <italic>MZsap130a</italic> hearts have large bulbus arteriosus <bold>(A, B)</bold> <italic>AB&#x2a;</italic> and <italic>MZsap130a</italic> adult SV hearts extracted at approximately 4&#x2013;6&#xa0;months post fertilization. <bold>(C)</bold> Quantification of ventricle area of unfixed hearts, pval is from a one ANOVA solid black bars are mean, dotted lines represent up and lower 25th percentiles. Each point represents a single heart, <italic>AB&#x2a;, n</italic> &#x3d; 14; <italic>MZsap130a</italic> NV, <italic>n</italic> &#x3d; 7; <italic>MZsap130a</italic> SV, <italic>n</italic> &#x3d; 12. <bold>(D)</bold> Quantification of BA area. of unfixed hearts, pval is from a one ANOVA solid black bars are mean, dotted lines represent up and lower 25th percentiles. Each point represents a single heart, <italic>n</italic> &#x3d; 10 for all groups. V, A, and BA are ventricle, atria, and bulbus arteriosus respectively, Scale bar 200&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g008.tif"/>
</fig>
</sec>
<sec id="s3-5">
<title>
<italic>sap130a</italic> genetically interacts with <italic>hdac1</italic> during SHF accretion</title>
<p>Zebrafish <italic>hdac1</italic> is required for ventricle formation (<xref ref-type="bibr" rid="B78">Song et al., 2019</xref>; <xref ref-type="bibr" rid="B11">Buhler et al., 2021</xref>). We explored the potential interaction of Sap130a and Hdac1 by analyzing heart development in <italic>MZsap130a;hdac1</italic>
<sup>
<italic>&#x2b;/b382</italic>
</sup> embryos. While <italic>hdac1</italic> homozygous mutants develop cardiac defects, heterozygous mutants are viable and show a similar proportion of SVs like in the <italic>MZsap130a</italic> mutants. An increase in SV phenotype was noted in <italic>MZsap130a;hdac1</italic>
<sup>
<italic>&#x2b;/b382</italic>
</sup> suggesting <italic>MZsap130a</italic> mutants are sensitized to <italic>hdac1</italic> gene dosage (<xref ref-type="fig" rid="F9">Figures 9A, C</xref>). These data revealed an association between <italic>hdac1</italic> heterozygous status and ventricle size and this increase in a <italic>MZsap130a</italic> background (<xref ref-type="fig" rid="F9">Figure 9C</xref>). This suggests that <italic>sap130a</italic> and <italic>hdac1</italic> genetically interact in zebrafish and function in the same complex like in mammals.</p>
<fig id="F9" position="float">
<label>FIGURE 9</label>
<caption>
<p>
<italic>sap130a</italic> shows an association with <italic>hdac1</italic> and <italic>MZsin3ab</italic> mutants have SVs <bold>(A)</bold> Representative image of <italic>Tg(myl7:EGFP)</italic> and <italic>MZsap130a;hdac1</italic>
<sup>
<italic>b382/&#x2b;</italic>
</sup>
<italic>;Tg</italic>(<italic>myl7:EGFP</italic>) NV and SVs at 48hpf. <bold>(B)</bold> Representative image of <italic>Tg(myl7:EGFP)</italic> and <italic>MZsin3ab</italic>;<italic>Tg(myl7:EGFP)</italic> NV and SVs at 48hpf. <bold>(C)</bold> Quantification of heart phenotype proportions in <italic>Tg(myl7:EGFP)</italic>, <italic>MZsap130a;Tg(myl7:EGFP)</italic>, <italic>hdac1</italic>
<sup>
<italic>b382/&#x2b;</italic>
</sup>
<italic>;Tg</italic>(<italic>myl7:EGFP</italic>) and <italic>MZsap130a;hdac1</italic>
<sup>
<italic>b382/&#x2b;</italic>
</sup>
<italic>;Tg</italic>(<italic>myl7:EGFP</italic>). The <italic>p</italic>-values are fisher&#x2019;s exact test. <bold>(D)</bold> Quantification of heart phenotype proportions in <italic>Tg(myl7:EGFP)</italic> and <italic>MZsin3ab;Tg(myl7:EGFP).</italic> The <italic>p</italic>-values are fisher&#x2019;s exact test. V and A are ventricle and atria, respectively. Scale bar 100&#xa0;&#x3bc;m.</p>
</caption>
<graphic xlink:href="fcell-11-1197109-g009.tif"/>
</fig>
<p>Both <italic>MZsap130a</italic> whole embryo and heart specific RNA-seq datasets revealed sarcomere genes to be down and cell cycle genes to be upregulated, similar to SIN3A knock-out and knock-down studies (<xref ref-type="sec" rid="s11">Supplementary Figure S12</xref>) (<xref ref-type="bibr" rid="B85">van Oevelen et al., 2008</xref>; <xref ref-type="bibr" rid="B84">van Oevelen et al., 2010</xref>; <xref ref-type="bibr" rid="B21">Dobi et al., 2014</xref>). For example, the cell cycle genes <italic>vrk1</italic>, <italic>e2f7</italic>, and <italic>haus6</italic> are increased in <italic>MZsap130a</italic> mutants, while we did not find evidence of expanded cardiac progenitors. These similarities in up and down DEGs point to the possibility that <italic>sap130a</italic> associates with <italic>sin3aa</italic> or <italic>sin3ab</italic> in zebrafish, similar to mammals. To further explore the importance of SIN3A in heart development, we generated <italic>MZsin3ab</italic> mutants using CRISPR/Cas9. The <italic>sin3ab pt36a</italic> allele generated disrupted amino acids 862&#x2013;867. In <italic>MZsin3ab</italic> mutants an in complete penetrant SV phenotype was observed in 44% (<xref ref-type="fig" rid="F9">Figures 9B, D</xref>). WISH data at 30hpf, revealed that <italic>ltbp3</italic> expression in <italic>MZsin3ab</italic> mutants was reduced, similar to the <italic>MZsap130a</italic> mutants in <xref ref-type="sec" rid="s11">Supplementary Figure S8</xref>. It is not surprising that the penetrance of the phenotype in <italic>MZsin3ab</italic> was also incomplete since both <italic>sin3aa</italic> and <italic>sin3b</italic> could compensate for the disruption of <italic>sin3ab</italic>. These data suggest that <italic>sin3ab</italic> is involved in ventricular development in zebrafish, a phenotype that is reminiscent of <italic>sap130a</italic> and <italic>hdac1</italic> mutants.</p>
</sec>
</sec>
<sec sec-type="discussion" id="s4">
<title>Discussion</title>
<p>In this study, we have revealed a role for <italic>sap130a</italic> in zebrafish cardiogenesis. We describe a null allele of <italic>sap130a</italic>, resulting in small ventricles through the delay and failure of SHF cells to migrate into the ventricle. Without <italic>sap130a</italic>, some of the SHF progenitors permanently become OFT cells. Transcriptome profiling of the <italic>MZsap130a</italic> embryos at 36hpf and hearts at 48hpf revealed that expression of sarcomere, conduction, and metabolism genes were dysregulated. This suggest that the CMs fail to terminally differentiate and properly function.</p>
<p>Our study reveals the consequence of disrupting members of the SIN3A complex, resulting in improper heart development. In the <italic>MZsap130a</italic> mutants, the main phenotype is a small ventricle leading to larger OFT and bulbus arteriosus in adulthood. Developmentally this arises from the failure of SHF progenitors to migrate into the growing ventricle. We come to this conclusion because the WISH data for <italic>nkx2.5</italic> and <italic>myh7</italic> showed no changes prior to the 24 hpf, indicating the FHF is intact. The phenotype arising at 36hpf is in line with observations showing the addition of SHF cells between 24 and 48hpf and with our lineage tracing experiments (<xref ref-type="bibr" rid="B30">Grimes et al., 2008</xref>; <xref ref-type="bibr" rid="B20">de Pater et al., 2009</xref>; <xref ref-type="bibr" rid="B32">Hami et al., 2011</xref>; <xref ref-type="bibr" rid="B52">Lazic and Scott, 2011</xref>; <xref ref-type="bibr" rid="B24">Felker et al., 2018</xref>). In the <italic>Ohia</italic> mouse mutant, the combination of <italic>PCDHA9</italic> and a <italic>SAP130</italic> mutations caused an HLHS etiology influencing the FHF structures. The prominent phenotype included a hypoplastic left ventricle and valve abnormalities in 11% of mouse embryos. In the zebrafish, the <italic>sap130a</italic> mutation is predicted to be a null mutant producing a hypoplastic ventricle in 48% of embryos. The difference seen between the mouse and zebrafish can be explained by the difference in the number of ventricle chambers, the specialized development of the mammalian OFT, and the changes seen during the evolution of this specialized pump from sea to land (<xref ref-type="bibr" rid="B43">Jensen et al., 2013</xref>). A recent study of <italic>SAP130</italic> pig CRISPR mutants show tricuspid dysplasia and atresia, highlighting the complex role of Sap130 in heart development across different species (<xref ref-type="bibr" rid="B26">Gabriel et al., 2021</xref>).</p>
<p>The catalytic unit of the SIN3A complex is comprised of class I HDACs, which deacetylate lysine residues to alter gene expression or protein function. The <italic>hdac1</italic> mutant, <italic>cardiac really gone (crg)</italic> exhibit decreased ventricular CMs by 36hpf (<xref ref-type="bibr" rid="B78">Song et al., 2019</xref>). This is also the timepoint <italic>MZsap130a</italic> mutants show decreased ventricular size. This suggests <italic>sap130a</italic> and <italic>hdac1</italic> could have overlapping functions during SHF development. Although SAP130 was shown to have HDAC-independent functions from <italic>in vitro</italic> studies (<xref ref-type="bibr" rid="B25">Fleischer et al., 2003</xref>), genetic studies suggest <italic>sap130a</italic> and <italic>hdac1</italic> interact for proper ventricular cardiomyocyte development. This supports a model in which Sap130a associates with the Sin3a/Hdac1 complex and/or an Hdac1-independent X-factor to regulate transcription (<xref ref-type="sec" rid="s11">Supplementary Figure S13</xref>).</p>
<p>Taken together this study support the importance of Sap130a/Sin3a/Hdac complex in zebrafish cardiogenesis. These observations build upon previous studies in zebrafish <italic>hdac1</italic> and reiterates the importance of context specific components for the Sin3a complex during cardiogenesis.</p>
</sec>
</body>
<back>
<sec sec-type="data-availability" id="s5">
<title>Data availability statement</title>
<p>The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: NCBI GEO under GSE228451.</p>
</sec>
<sec id="s6">
<title>Ethics statement</title>
<p>The animal study was approved by Institutional Animal Care and Use Committee (IACUC) at the University of Pittsburgh. The study was conducted in accordance with the local legislation and institutional requirements.</p>
</sec>
<sec id="s7">
<title>Author contributions</title>
<p>RD, CL, and MT designed research; RD, RF-R, DF, and JS performed research; RD and MT analyzed data CL and MT responsible for funding. RD wrote the manuscript. All authors contributed to the article and approved the submitted version.</p>
</sec>
<sec id="s8">
<title>Funding</title>
<p>This research was supported by funding from the National Institutes of Health R01HL142788 to MT and CL, and R01HL156398 to MT This research was supported in part by the University of Pittsburgh Center for Research Computing through the resources provided. Open Access funding provided by University of Pittsburgh. This research was supported in part by the University of Pittsburgh Center for Research Computing, RRID: SCR_022735, through the resources provided. Specifically, this work used the HTC cluster, which is supported by NIH award number S10OD028483. This project used the University of Pittsburgh HSCRF Genomics Research Core, RRID: SCR_018301, for RNA-seq experiments. RNA-seq data were aligned with CLC genomics Workbench Version 20 (QIAGEN), licensed through the Molecular Biology Information Service of the Health Sciences Library System, University of Pittsburgh.</p>
</sec>
<ack>
<p>We are grateful to members of the MT lab, Donghun JS, Elizabeth Rochon, and Neil Hukriede for reading the manuscript and experimental suggestions. Tim Feinstein, who built the light sheet scope and mounting system. Manush Saydmohammed and Shoulin Li for initial work on <italic>MZsap130a</italic> mutant and the <italic>Tg(myl7:memGFP)</italic> line.</p>
</ack>
<sec sec-type="COI-statement" id="s9">
<title>Conflict of interest</title>
<p>The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p>
</sec>
<sec sec-type="disclaimer" id="s10">
<title>Publisher&#x2019;s note</title>
<p>All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.</p>
</sec>
<sec id="s11">
<title>Supplementary material</title>
<p>The Supplementary Material for this article can be found online at: <ext-link ext-link-type="uri" xlink:href="https://www.frontiersin.org/articles/10.3389/fcell.2023.1197109/full#supplementary-material">https://www.frontiersin.org/articles/10.3389/fcell.2023.1197109/full&#x23;supplementary-material</ext-link>
</p>
<supplementary-material xlink:href="Table2.XLSX" id="SM1" mimetype="application/XLSX" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Video2.AVI" id="SM2" mimetype="application/AVI" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Video3.AVI" id="SM3" mimetype="application/AVI" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Table3.XLSX" id="SM4" mimetype="application/XLSX" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Table6.XLSX" id="SM5" mimetype="application/XLSX" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="DataSheet1.PDF" id="SM6" mimetype="application/PDF" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Table4.XLSX" id="SM7" mimetype="application/XLSX" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Video5.AVI" id="SM8" mimetype="application/AVI" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Table1.XLSX" id="SM9" mimetype="application/XLSX" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Table5.XLSX" id="SM10" mimetype="application/XLSX" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Video1.AVI" id="SM11" mimetype="application/AVI" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Video4.AVI" id="SM12" mimetype="application/AVI" xmlns:xlink="http://www.w3.org/1999/xlink"/>
</sec>
<ref-list>
<title>References</title>
<ref id="B1">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Adams</surname>
<given-names>G. E.</given-names>
</name>
<name>
<surname>Chandru</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Cowley</surname>
<given-names>S. M.</given-names>
</name>
</person-group> (<year>2018</year>). <article-title>Co-Repressor, co-activator and general transcription factor: the many faces of the Sin3 histone deacetylase (HDAC) complex</article-title>. <source>Biochem. J.</source> <volume>475</volume> (<issue>24</issue>), <fpage>3921</fpage>&#x2013;<lpage>3932</lpage>. <pub-id pub-id-type="doi">10.1042/BCJ20170314</pub-id>
</citation>
</ref>
<ref id="B2">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Alfert</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Moreno</surname>
<given-names>N.</given-names>
</name>
<name>
<surname>Kerl</surname>
<given-names>K.</given-names>
</name>
</person-group> (<year>2019</year>). <article-title>The BAF complex in development and disease</article-title>. <source>Epigenetics Chromatin</source> <volume>12</volume> (<issue>1</issue>), <fpage>19</fpage>. <pub-id pub-id-type="doi">10.1186/s13072-019-0264-y</pub-id>
</citation>
</ref>
<ref id="B3">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Alsop</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Vijayan</surname>
<given-names>M.</given-names>
</name>
</person-group> (<year>2009</year>). <article-title>The zebrafish stress axis: molecular fallout from the teleost-specific genome duplication event</article-title>. <source>Gen. Comp. Endocrinol.</source> <volume>161</volume> (<issue>1</issue>), <fpage>62</fpage>&#x2013;<lpage>66</lpage>. <pub-id pub-id-type="doi">10.1016/j.ygcen.2008.09.011</pub-id>
</citation>
</ref>
<ref id="B4">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Auman</surname>
<given-names>H. J.</given-names>
</name>
<name>
<surname>Fernandes</surname>
<given-names>I. H.</given-names>
</name>
<name>
<surname>Berrios-Otero</surname>
<given-names>C. A.</given-names>
</name>
<name>
<surname>Colombo</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Yelon</surname>
<given-names>D.</given-names>
</name>
</person-group> (<year>2023</year>). <article-title>Zebrafish smarcc1a mutants reveal requirements for BAF chromatin remodeling complexes in distinguishing the atrioventricular canal from the cardiac chambers</article-title>. <source>Dev. Dyn</source>. <pub-id pub-id-type="doi">10.1002/dvdy.595</pub-id>
</citation>
</ref>
<ref id="B5">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Barron</surname>
<given-names>D. J.</given-names>
</name>
<name>
<surname>Kilby</surname>
<given-names>M. D.</given-names>
</name>
<name>
<surname>Davies</surname>
<given-names>B.</given-names>
</name>
<name>
<surname>Wright</surname>
<given-names>J. G.</given-names>
</name>
<name>
<surname>Jones</surname>
<given-names>T. J.</given-names>
</name>
<name>
<surname>Brawn</surname>
<given-names>W. J.</given-names>
</name>
</person-group> (<year>2009</year>). <article-title>Hypoplastic left heart syndrome</article-title>. <source>Lancet</source> <volume>374</volume> (<issue>9689</issue>), <fpage>551</fpage>&#x2013;<lpage>564</lpage>. <pub-id pub-id-type="doi">10.1016/S0140-6736(09)60563-8</pub-id>
</citation>
</ref>
<ref id="B6">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Berezin</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Glaser</surname>
<given-names>F.</given-names>
</name>
<name>
<surname>Rosenberg</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Paz</surname>
<given-names>I.</given-names>
</name>
<name>
<surname>Pupko</surname>
<given-names>T.</given-names>
</name>
<name>
<surname>Fariselli</surname>
<given-names>P.</given-names>
</name>
<etal/>
</person-group> (<year>2004</year>). <article-title>ConSeq: the identification of functionally and structurally important residues in protein sequences</article-title>. <source>Bioinformatics</source> <volume>20</volume> (<issue>8</issue>), <fpage>1322</fpage>&#x2013;<lpage>1324</lpage>. <pub-id pub-id-type="doi">10.1093/bioinformatics/bth070</pub-id>
</citation>
</ref>
<ref id="B7">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Bisserier</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Mathiyalagan</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Zhang</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Elmastour</surname>
<given-names>F.</given-names>
</name>
<name>
<surname>Dorfmuller</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Humbert</surname>
<given-names>M.</given-names>
</name>
<etal/>
</person-group> (<year>2021</year>). <article-title>Regulation of the methylation and expression levels of the BMPR2 gene by SIN3a as a novel therapeutic mechanism in pulmonary arterial hypertension</article-title>. <source>Circulation</source> <volume>144</volume> (<issue>1</issue>), <fpage>52</fpage>&#x2013;<lpage>73</lpage>. <pub-id pub-id-type="doi">10.1161/CIRCULATIONAHA.120.047978</pub-id>
</citation>
</ref>
<ref id="B8">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Bodenhofer</surname>
<given-names>U.</given-names>
</name>
<name>
<surname>Bonatesta</surname>
<given-names>E.</given-names>
</name>
<name>
<surname>Horejs-Kainrath</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Hochreiter</surname>
<given-names>S.</given-names>
</name>
</person-group> (<year>2015</year>). <article-title>msa: an R package for multiple sequence alignment</article-title>. <source>Bioinformatics</source> <volume>31</volume> (<issue>24</issue>), <fpage>3997</fpage>&#x2013;<lpage>3999</lpage>. <pub-id pub-id-type="doi">10.1093/bioinformatics/btv494</pub-id>
</citation>
</ref>
<ref id="B9">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Brennan</surname>
<given-names>P.</given-names>
</name>
</person-group> (<year>2018</year>). <article-title>drawProteins: a Bioconductor/R package for reproducible and programmatic generation of protein schematics</article-title>. <source>F1000Res</source> <volume>7</volume>, <fpage>1105</fpage>. <pub-id pub-id-type="doi">10.12688/f1000research.14541.1</pub-id>
</citation>
</ref>
<ref id="B10">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Brown</surname>
<given-names>M. A.</given-names>
</name>
<name>
<surname>Sims</surname>
<given-names>R. J.</given-names>
</name>
<name>
<surname>Gottlieb</surname>
<given-names>P. D.</given-names>
</name>
<name>
<surname>Tucker</surname>
<given-names>P. W.</given-names>
</name>
</person-group> (<year>2006</year>). <article-title>Identification and characterization of Smyd2: A split SET/MYND domain-containing histone H3 lysine 36-specific methyltransferase that interacts with the Sin3 histone deacetylase complex</article-title>. <source>Mol. Cancer</source> <volume>5</volume>, <fpage>26</fpage>. <pub-id pub-id-type="doi">10.1186/1476-4598-5-26</pub-id>
</citation>
</ref>
<ref id="B11">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Buhler</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Gahr</surname>
<given-names>B. M.</given-names>
</name>
<name>
<surname>Park</surname>
<given-names>D. D.</given-names>
</name>
<name>
<surname>Bertozzi</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Boos</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Dalvoy</surname>
<given-names>M.</given-names>
</name>
<etal/>
</person-group> (<year>2021</year>). <article-title>Histone deacetylase 1 controls cardiomyocyte proliferation during embryonic heart development and cardiac regeneration in zebrafish</article-title>. <source>PLoS Genet.</source> <volume>17</volume> (<issue>11</issue>), <fpage>e1009890</fpage>. <pub-id pub-id-type="doi">10.1371/journal.pgen.1009890</pub-id>
</citation>
</ref>
<ref id="B12">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Carrozza</surname>
<given-names>M. J.</given-names>
</name>
<name>
<surname>Li</surname>
<given-names>B.</given-names>
</name>
<name>
<surname>Florens</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Suganuma</surname>
<given-names>T.</given-names>
</name>
<name>
<surname>Swanson</surname>
<given-names>S. K.</given-names>
</name>
<name>
<surname>Lee</surname>
<given-names>K. K.</given-names>
</name>
<etal/>
</person-group> (<year>2005</year>). <article-title>Histone H3 methylation by Set2 directs deacetylation of coding regions by Rpd3S to suppress spurious intragenic transcription</article-title>. <source>Cell</source> <volume>123</volume> (<issue>4</issue>), <fpage>581</fpage>&#x2013;<lpage>592</lpage>. <pub-id pub-id-type="doi">10.1016/j.cell.2005.10.023</pub-id>
</citation>
</ref>
<ref id="B13">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Charif</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Thioulouse</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Lobry</surname>
<given-names>J. R.</given-names>
</name>
<name>
<surname>Perriere</surname>
<given-names>G.</given-names>
</name>
</person-group> (<year>2005</year>). <article-title>Online synonymous codon usage analyses with the ade4 and seqinR packages</article-title>. <source>Bioinformatics</source> <volume>21</volume> (<issue>4</issue>), <fpage>545</fpage>&#x2013;<lpage>547</lpage>. <pub-id pub-id-type="doi">10.1093/bioinformatics/bti037</pub-id>
</citation>
</ref>
<ref id="B14">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Chen</surname>
<given-names>Q.</given-names>
</name>
<name>
<surname>Chen</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Jian</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Li</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Zhang</surname>
<given-names>X.</given-names>
</name>
</person-group> (<year>2022</year>). <article-title>The mechanism behind BAF60c in myocardial metabolism in rats with heart failure is through the PGC1&#x3b1;-PPAR&#x3b1;-mTOR signaling pathway</article-title>. <source>Biochem. Cell Biol.</source> <volume>100</volume> (<issue>2</issue>), <fpage>93</fpage>&#x2013;<lpage>103</lpage>. <pub-id pub-id-type="doi">10.1139/bcb-2019-0450</pub-id>
</citation>
</ref>
<ref id="B15">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Chi</surname>
<given-names>N. C.</given-names>
</name>
<name>
<surname>Bussen</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Brand-Arzamendi</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>Ding</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Olgin</surname>
<given-names>J. E.</given-names>
</name>
<name>
<surname>Shaw</surname>
<given-names>R. M.</given-names>
</name>
<etal/>
</person-group> (<year>2010</year>). <article-title>Cardiac conduction is required to preserve cardiac chamber morphology</article-title>. <source>Proc. Natl. Acad. Sci. U. S. A.</source> <volume>107</volume> (<issue>33</issue>), <fpage>14662</fpage>&#x2013;<lpage>14667</lpage>. <pub-id pub-id-type="doi">10.1073/pnas.0909432107</pub-id>
</citation>
</ref>
<ref id="B16">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Ching</surname>
<given-names>Y. H.</given-names>
</name>
<name>
<surname>Ghosh</surname>
<given-names>T. K.</given-names>
</name>
<name>
<surname>Cross</surname>
<given-names>S. J.</given-names>
</name>
<name>
<surname>Packham</surname>
<given-names>E. A.</given-names>
</name>
<name>
<surname>Honeyman</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Loughna</surname>
<given-names>S.</given-names>
</name>
<etal/>
</person-group> (<year>2005</year>). <article-title>Mutation in myosin heavy chain 6 causes atrial septal defect</article-title>. <source>Nat. Genet.</source> <volume>37</volume> (<issue>4</issue>), <fpage>423</fpage>&#x2013;<lpage>428</lpage>. <pub-id pub-id-type="doi">10.1038/ng1526</pub-id>
</citation>
</ref>
<ref id="B17">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Connor</surname>
<given-names>J. A.</given-names>
</name>
<name>
<surname>Thiagarajan</surname>
<given-names>R.</given-names>
</name>
</person-group> (<year>2007</year>). <article-title>Hypoplastic left heart syndrome</article-title>. <source>Orphanet J. Rare Dis.</source> <volume>2</volume>, <fpage>23</fpage>. <pub-id pub-id-type="doi">10.1186/1750-1172-2-23</pub-id>
</citation>
</ref>
<ref id="B18">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Costain</surname>
<given-names>G.</given-names>
</name>
<name>
<surname>Silversides</surname>
<given-names>C. K.</given-names>
</name>
<name>
<surname>Bassett</surname>
<given-names>A. S.</given-names>
</name>
</person-group> (<year>2016</year>). <article-title>The importance of copy number variation in congenital heart disease</article-title>. <source>NPJ Genom Med.</source> <volume>1</volume>, <fpage>16031</fpage>. <pub-id pub-id-type="doi">10.1038/npjgenmed.2016.31</pub-id>
</citation>
</ref>
<ref id="B19">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Dannenberg</surname>
<given-names>J. H.</given-names>
</name>
<name>
<surname>David</surname>
<given-names>G.</given-names>
</name>
<name>
<surname>Zhong</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>van der Torre</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Wong</surname>
<given-names>W. H.</given-names>
</name>
<name>
<surname>Depinho</surname>
<given-names>R. A.</given-names>
</name>
</person-group> (<year>2005</year>). <article-title>mSin3A corepressor regulates diverse transcriptional networks governing normal and neoplastic growth and survival</article-title>. <source>Genes Dev.</source> <volume>19</volume> (<issue>13</issue>), <fpage>1581</fpage>&#x2013;<lpage>1595</lpage>. <pub-id pub-id-type="doi">10.1101/gad.1286905</pub-id>
</citation>
</ref>
<ref id="B20">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>de Pater</surname>
<given-names>E.</given-names>
</name>
<name>
<surname>Clijsters</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Marques</surname>
<given-names>S. R.</given-names>
</name>
<name>
<surname>Lin</surname>
<given-names>Y. F.</given-names>
</name>
<name>
<surname>Garavito-Aguilar</surname>
<given-names>Z. V.</given-names>
</name>
<name>
<surname>Yelon</surname>
<given-names>D.</given-names>
</name>
<etal/>
</person-group> (<year>2009</year>). <article-title>Distinct phases of cardiomyocyte differentiation regulate growth of the zebrafish heart</article-title>. <source>Development</source> <volume>136</volume> (<issue>10</issue>), <fpage>1633</fpage>&#x2013;<lpage>1641</lpage>. <pub-id pub-id-type="doi">10.1242/dev.030924</pub-id>
</citation>
</ref>
<ref id="B21">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Dobi</surname>
<given-names>K. C.</given-names>
</name>
<name>
<surname>Halfon</surname>
<given-names>M. S.</given-names>
</name>
<name>
<surname>Baylies</surname>
<given-names>M. K.</given-names>
</name>
</person-group> (<year>2014</year>). <article-title>Whole-genome analysis of muscle founder cells implicates the chromatin regulator Sin3A in muscle identity</article-title>. <source>Cell Rep.</source> <volume>8</volume> (<issue>3</issue>), <fpage>858</fpage>&#x2013;<lpage>870</lpage>. <pub-id pub-id-type="doi">10.1016/j.celrep.2014.07.005</pub-id>
</citation>
</ref>
<ref id="B22">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Donlin</surname>
<given-names>L. T.</given-names>
</name>
<name>
<surname>Andresen</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Just</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Rudensky</surname>
<given-names>E.</given-names>
</name>
<name>
<surname>Pappas</surname>
<given-names>C. T.</given-names>
</name>
<name>
<surname>Kruger</surname>
<given-names>M.</given-names>
</name>
<etal/>
</person-group> (<year>2012</year>). <article-title>Smyd2 controls cytoplasmic lysine methylation of Hsp90 and myofilament organization</article-title>. <source>Genes Dev.</source> <volume>26</volume> (<issue>2</issue>), <fpage>114</fpage>&#x2013;<lpage>119</lpage>. <pub-id pub-id-type="doi">10.1101/gad.177758.111</pub-id>
</citation>
</ref>
<ref id="B23">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Fahed</surname>
<given-names>A. C.</given-names>
</name>
<name>
<surname>Gelb</surname>
<given-names>B. D.</given-names>
</name>
<name>
<surname>Seidman</surname>
<given-names>J. G.</given-names>
</name>
<name>
<surname>Seidman</surname>
<given-names>C. E.</given-names>
</name>
</person-group> (<year>2013</year>). <article-title>Genetics of congenital heart disease: the glass half empty</article-title>. <source>Circ. Res.</source> <volume>112</volume> (<issue>4</issue>), <fpage>707</fpage>&#x2013;<lpage>720</lpage>. <pub-id pub-id-type="doi">10.1161/CIRCRESAHA.112.300853</pub-id>
</citation>
</ref>
<ref id="B24">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Felker</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Prummel</surname>
<given-names>K. D.</given-names>
</name>
<name>
<surname>Merks</surname>
<given-names>A. M.</given-names>
</name>
<name>
<surname>Mickoleit</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Brombacher</surname>
<given-names>E. C.</given-names>
</name>
<name>
<surname>Huisken</surname>
<given-names>J.</given-names>
</name>
<etal/>
</person-group> (<year>2018</year>). <article-title>Continuous addition of progenitors forms the cardiac ventricle in zebrafish</article-title>. <source>Nat. Commun.</source> <volume>9</volume> (<issue>1</issue>), <fpage>2001</fpage>. <pub-id pub-id-type="doi">10.1038/s41467-018-04402-6</pub-id>
</citation>
</ref>
<ref id="B25">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Fleischer</surname>
<given-names>T. C.</given-names>
</name>
<name>
<surname>Yun</surname>
<given-names>U. J.</given-names>
</name>
<name>
<surname>Ayer</surname>
<given-names>D. E.</given-names>
</name>
</person-group> (<year>2003</year>). <article-title>Identification and characterization of three new components of the mSin3A corepressor complex</article-title>. <source>Mol. Cell Biol.</source> <volume>23</volume> (<issue>10</issue>), <fpage>3456</fpage>&#x2013;<lpage>3467</lpage>. <pub-id pub-id-type="doi">10.1128/mcb.23.10.3456-3467.2003</pub-id>
</citation>
</ref>
<ref id="B26">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Gabriel</surname>
<given-names>G. C.</given-names>
</name>
<name>
<surname>Devine</surname>
<given-names>W.</given-names>
</name>
<name>
<surname>Redel</surname>
<given-names>B. K.</given-names>
</name>
<name>
<surname>Whitworth</surname>
<given-names>K. M.</given-names>
</name>
<name>
<surname>Samuel</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Spate</surname>
<given-names>L. D.</given-names>
</name>
<etal/>
</person-group> (<year>2021</year>). <article-title>Cardiovascular development and congenital heart disease modeling in the pig</article-title>. <source>J. Am. Heart Assoc.</source> <volume>10</volume> (<issue>14</issue>), <fpage>e021631</fpage>. <pub-id pub-id-type="doi">10.1161/JAHA.121.021631</pub-id>
</citation>
</ref>
<ref id="B27">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Gagnon</surname>
<given-names>J. A.</given-names>
</name>
<name>
<surname>Valen</surname>
<given-names>E.</given-names>
</name>
<name>
<surname>Thyme</surname>
<given-names>S. B.</given-names>
</name>
<name>
<surname>Huang</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Akhmetova</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Pauli</surname>
<given-names>A.</given-names>
</name>
<etal/>
</person-group> (<year>2014</year>). <article-title>Efficient mutagenesis by Cas9 protein-mediated oligonucleotide insertion and large-scale assessment of single-guide RNAs</article-title>. <source>PLoS One</source> <volume>9</volume> (<issue>5</issue>), <fpage>e98186</fpage>. <pub-id pub-id-type="doi">10.1371/journal.pone.0098186</pub-id>
</citation>
</ref>
<ref id="B28">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Girstmair</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Zakrzewski</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Lapraz</surname>
<given-names>F.</given-names>
</name>
<name>
<surname>Handberg-Thorsager</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Tomancak</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Pitrone</surname>
<given-names>P. G.</given-names>
</name>
<etal/>
</person-group> (<year>2016</year>). <article-title>Light-sheet microscopy for everyone? Experience of building an OpenSPIM to study flatworm development</article-title>. <source>BMC Dev. Biol.</source> <volume>16</volume> (<issue>1</issue>), <fpage>22</fpage>. <pub-id pub-id-type="doi">10.1186/s12861-016-0122-0</pub-id>
</citation>
</ref>
<ref id="B29">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Granados-Riveron</surname>
<given-names>J. T.</given-names>
</name>
<name>
<surname>Ghosh</surname>
<given-names>T. K.</given-names>
</name>
<name>
<surname>Pope</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Bu&#x27;Lock</surname>
<given-names>F.</given-names>
</name>
<name>
<surname>Thornborough</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Eason</surname>
<given-names>J.</given-names>
</name>
<etal/>
</person-group> (<year>2010</year>). <article-title>Alpha-cardiac myosin heavy chain (MYH6) mutations affecting myofibril formation are associated with congenital heart defects</article-title>. <source>Hum. Mol. Genet.</source> <volume>19</volume> (<issue>20</issue>), <fpage>4007</fpage>&#x2013;<lpage>4016</lpage>. <pub-id pub-id-type="doi">10.1093/hmg/ddq315</pub-id>
</citation>
</ref>
<ref id="B30">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Grimes</surname>
<given-names>A. C.</given-names>
</name>
<name>
<surname>Erwin</surname>
<given-names>K. N.</given-names>
</name>
<name>
<surname>Stadt</surname>
<given-names>H. A.</given-names>
</name>
<name>
<surname>Hunter</surname>
<given-names>G. L.</given-names>
</name>
<name>
<surname>Gefroh</surname>
<given-names>H. A.</given-names>
</name>
<name>
<surname>Tsai</surname>
<given-names>H. J.</given-names>
</name>
<etal/>
</person-group> (<year>2008</year>). <article-title>PCB126 exposure disrupts zebrafish ventricular and branchial but not early neural crest development</article-title>. <source>Toxicol. Sci.</source> <volume>106</volume> (<issue>1</issue>), <fpage>193</fpage>&#x2013;<lpage>205</lpage>. <pub-id pub-id-type="doi">10.1093/toxsci/kfn154</pub-id>
</citation>
</ref>
<ref id="B31">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Guner-Ataman</surname>
<given-names>B.</given-names>
</name>
<name>
<surname>Paffett-Lugassy</surname>
<given-names>N.</given-names>
</name>
<name>
<surname>Adams</surname>
<given-names>M. S.</given-names>
</name>
<name>
<surname>Nevis</surname>
<given-names>K. R.</given-names>
</name>
<name>
<surname>Jahangiri</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Obregon</surname>
<given-names>P.</given-names>
</name>
<etal/>
</person-group> (<year>2013</year>). <article-title>Zebrafish second heart field development relies on progenitor specification in anterior lateral plate mesoderm and nkx2.5 function</article-title>. <source>Development</source> <volume>140</volume> (<issue>6</issue>), <fpage>1353</fpage>&#x2013;<lpage>1363</lpage>. <pub-id pub-id-type="doi">10.1242/dev.088351</pub-id>
</citation>
</ref>
<ref id="B32">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Hami</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Grimes</surname>
<given-names>A. C.</given-names>
</name>
<name>
<surname>Tsai</surname>
<given-names>H. J.</given-names>
</name>
<name>
<surname>Kirby</surname>
<given-names>M. L.</given-names>
</name>
</person-group> (<year>2011</year>). <article-title>Zebrafish cardiac development requires a conserved secondary heart field</article-title>. <source>Development</source> <volume>138</volume> (<issue>11</issue>), <fpage>2389</fpage>&#x2013;<lpage>2398</lpage>. <pub-id pub-id-type="doi">10.1242/dev.061473</pub-id>
</citation>
</ref>
<ref id="B33">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Han</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Hang</surname>
<given-names>C. T.</given-names>
</name>
<name>
<surname>Yang</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Chang</surname>
<given-names>C. P.</given-names>
</name>
</person-group> (<year>2011</year>). <article-title>Chromatin remodeling in cardiovascular development and physiology</article-title>. <source>Circ. Res.</source> <volume>108</volume> (<issue>3</issue>), <fpage>378</fpage>&#x2013;<lpage>396</lpage>. <pub-id pub-id-type="doi">10.1161/CIRCRESAHA.110.224287</pub-id>
</citation>
</ref>
<ref id="B34">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Hang</surname>
<given-names>C. T.</given-names>
</name>
<name>
<surname>Yang</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Han</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Cheng</surname>
<given-names>H. L.</given-names>
</name>
<name>
<surname>Shang</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Ashley</surname>
<given-names>E.</given-names>
</name>
<etal/>
</person-group> (<year>2010</year>). <article-title>Chromatin regulation by Brg1 underlies heart muscle development and disease</article-title>. <source>Nature</source> <volume>466</volume> (<issue>7302</issue>), <fpage>62</fpage>&#x2013;<lpage>67</lpage>. <pub-id pub-id-type="doi">10.1038/nature09130</pub-id>
</citation>
</ref>
<ref id="B35">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Hargreaves</surname>
<given-names>D. C.</given-names>
</name>
<name>
<surname>Crabtree</surname>
<given-names>G. R.</given-names>
</name>
</person-group> (<year>2011</year>). <article-title>ATP-Dependent chromatin remodeling: genetics, genomics and mechanisms</article-title>. <source>Cell Res.</source> <volume>21</volume> (<issue>3</issue>), <fpage>396</fpage>&#x2013;<lpage>420</lpage>. <pub-id pub-id-type="doi">10.1038/cr.2011.32</pub-id>
</citation>
</ref>
<ref id="B36">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Haverinen</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Hassinen</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Dash</surname>
<given-names>S. N.</given-names>
</name>
<name>
<surname>Vornanen</surname>
<given-names>M.</given-names>
</name>
</person-group> (<year>2018</year>). <article-title>Expression of calcium channel transcripts in the zebrafish heart: dominance of T-type channels</article-title>. <source>J. Exp. Biol.</source> <volume>221</volume> (<issue>10</issue>), <fpage>jeb179226</fpage>. <pub-id pub-id-type="doi">10.1242/jeb.179226</pub-id>
</citation>
</ref>
<ref id="B37">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Hota</surname>
<given-names>S. K.</given-names>
</name>
<name>
<surname>Johnson</surname>
<given-names>J. R.</given-names>
</name>
<name>
<surname>Verschueren</surname>
<given-names>E.</given-names>
</name>
<name>
<surname>Thomas</surname>
<given-names>R.</given-names>
</name>
<name>
<surname>Blotnick</surname>
<given-names>A. M.</given-names>
</name>
<name>
<surname>Zhu</surname>
<given-names>Y.</given-names>
</name>
<etal/>
</person-group> (<year>2019</year>). <article-title>Dynamic BAF chromatin remodeling complex subunit inclusion promotes temporally distinct gene expression programs in cardiogenesis</article-title>. <source>Development</source> <volume>146</volume> (<issue>19</issue>), <fpage>dev174086</fpage>. <pub-id pub-id-type="doi">10.1242/dev.174086</pub-id>
</citation>
</ref>
<ref id="B38">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Huang</surname>
<given-names>C. J.</given-names>
</name>
<name>
<surname>Tu</surname>
<given-names>C. T.</given-names>
</name>
<name>
<surname>Hsiao</surname>
<given-names>C. D.</given-names>
</name>
<name>
<surname>Hsieh</surname>
<given-names>F. J.</given-names>
</name>
<name>
<surname>Tsai</surname>
<given-names>H. J.</given-names>
</name>
</person-group> (<year>2003</year>). <article-title>Germ-line transmission of a myocardium-specific GFP transgene reveals critical regulatory elements in the cardiac myosin light chain 2 promoter of zebrafish</article-title>. <source>Dev. Dyn.</source> <volume>228</volume> (<issue>1</issue>), <fpage>30</fpage>&#x2013;<lpage>40</lpage>. <pub-id pub-id-type="doi">10.1002/dvdy.10356</pub-id>
</citation>
</ref>
<ref id="B39">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Huang</surname>
<given-names>H. T.</given-names>
</name>
<name>
<surname>Kathrein</surname>
<given-names>K. L.</given-names>
</name>
<name>
<surname>Barton</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Gitlin</surname>
<given-names>Z.</given-names>
</name>
<name>
<surname>Huang</surname>
<given-names>Y. H.</given-names>
</name>
<name>
<surname>Ward</surname>
<given-names>T. P.</given-names>
</name>
<etal/>
</person-group> (<year>2013</year>). <article-title>A network of epigenetic regulators guides developmental haematopoiesis <italic>in vivo</italic>
</article-title>. <source>Nat. Cell Biol.</source> <volume>15</volume> (<issue>12</issue>), <fpage>1516</fpage>&#x2013;<lpage>1525</lpage>. <pub-id pub-id-type="doi">10.1038/ncb2870</pub-id>
</citation>
</ref>
<ref id="B40">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Huang</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Akerberg</surname>
<given-names>A. A.</given-names>
</name>
<name>
<surname>Zhang</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Yoon</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Joshi</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Hallinan</surname>
<given-names>C.</given-names>
</name>
<etal/>
</person-group> (<year>2022</year>). <article-title>Intrinsic myocardial defects underlie an Rbfox-deficient zebrafish model of hypoplastic left heart syndrome</article-title>. <source>Nat. Commun.</source> <volume>13</volume> (<issue>1</issue>), <fpage>5877</fpage>. <pub-id pub-id-type="doi">10.1038/s41467-022-32982-x</pub-id>
</citation>
</ref>
<ref id="B41">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Ignatius</surname>
<given-names>M. S.</given-names>
</name>
<name>
<surname>Unal Eroglu</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Malireddy</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Gallagher</surname>
<given-names>G.</given-names>
</name>
<name>
<surname>Nambiar</surname>
<given-names>R. M.</given-names>
</name>
<name>
<surname>Henion</surname>
<given-names>P. D.</given-names>
</name>
</person-group> (<year>2013</year>). <article-title>Distinct functional and temporal requirements for zebrafish Hdac1 during neural crest-derived craniofacial and peripheral neuron development</article-title>. <source>PLoS One</source> <volume>8</volume> (<issue>5</issue>), <fpage>e63218</fpage>. <pub-id pub-id-type="doi">10.1371/journal.pone.0063218</pub-id>
</citation>
</ref>
<ref id="B42">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Itou</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Oishi</surname>
<given-names>I.</given-names>
</name>
<name>
<surname>Kawakami</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Glass</surname>
<given-names>T. J.</given-names>
</name>
<name>
<surname>Richter</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Johnson</surname>
<given-names>A.</given-names>
</name>
<etal/>
</person-group> (<year>2012</year>). <article-title>Migration of cardiomyocytes is essential for heart regeneration in zebrafish</article-title>. <source>Development</source> <volume>139</volume> (<issue>22</issue>), <fpage>4133</fpage>&#x2013;<lpage>4142</lpage>. <pub-id pub-id-type="doi">10.1242/dev.079756</pub-id>
</citation>
</ref>
<ref id="B43">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Jensen</surname>
<given-names>B.</given-names>
</name>
<name>
<surname>Wang</surname>
<given-names>T.</given-names>
</name>
<name>
<surname>Christoffels</surname>
<given-names>V. M.</given-names>
</name>
<name>
<surname>Moorman</surname>
<given-names>A. F.</given-names>
</name>
</person-group> (<year>2013</year>). <article-title>Evolution and development of the building plan of the vertebrate heart</article-title>. <source>Biochim. Biophys. Acta</source> <volume>1833</volume> (<issue>4</issue>), <fpage>783</fpage>&#x2013;<lpage>794</lpage>. <pub-id pub-id-type="doi">10.1016/j.bbamcr.2012.10.004</pub-id>
</citation>
</ref>
<ref id="B44">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Jiang</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Jiang</surname>
<given-names>Z.</given-names>
</name>
<name>
<surname>Lu</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Wang</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Liang</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Zhang</surname>
<given-names>J.</given-names>
</name>
<etal/>
</person-group> (<year>2019</year>). <article-title>Migrasomes provide regional cues for organ morphogenesis during zebrafish gastrulation</article-title>. <source>Nat. Cell Biol.</source> <volume>21</volume> (<issue>8</issue>), <fpage>966</fpage>&#x2013;<lpage>977</lpage>. <pub-id pub-id-type="doi">10.1038/s41556-019-0358-6</pub-id>
</citation>
</ref>
<ref id="B45">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Jing</surname>
<given-names>L.</given-names>
</name>
</person-group> (<year>2012</year>). <article-title>Zebrafish embryo DNA preparation</article-title>. <source>Bio-protocol</source> <volume>2</volume>, <fpage>e184</fpage>. <pub-id pub-id-type="doi">10.21769/bioprotoc.184</pub-id>
</citation>
</ref>
<ref id="B46">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Jones</surname>
<given-names>W. K.</given-names>
</name>
<name>
<surname>Grupp</surname>
<given-names>I. L.</given-names>
</name>
<name>
<surname>Doetschman</surname>
<given-names>T.</given-names>
</name>
<name>
<surname>Grupp</surname>
<given-names>G.</given-names>
</name>
<name>
<surname>Osinska</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Hewett</surname>
<given-names>T. E.</given-names>
</name>
<etal/>
</person-group> (<year>1996</year>). <article-title>Ablation of the murine alpha myosin heavy chain gene leads to dosage effects and functional deficits in the heart</article-title>. <source>J. Clin. Invest.</source> <volume>98</volume> (<issue>8</issue>), <fpage>1906</fpage>&#x2013;<lpage>1917</lpage>. <pub-id pub-id-type="doi">10.1172/JCI118992</pub-id>
</citation>
</ref>
<ref id="B47">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Joshi</surname>
<given-names>A. A.</given-names>
</name>
<name>
<surname>Struhl</surname>
<given-names>K.</given-names>
</name>
</person-group> (<year>2005</year>). <article-title>Eaf3 chromodomain interaction with methylated H3-K36 links histone deacetylation to Pol II elongation</article-title>. <source>Mol. Cell</source> <volume>20</volume> (<issue>6</issue>), <fpage>971</fpage>&#x2013;<lpage>978</lpage>. <pub-id pub-id-type="doi">10.1016/j.molcel.2005.11.021</pub-id>
</citation>
</ref>
<ref id="B48">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Kadamb</surname>
<given-names>R.</given-names>
</name>
<name>
<surname>Mittal</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Bansal</surname>
<given-names>N.</given-names>
</name>
<name>
<surname>Batra</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Saluja</surname>
<given-names>D.</given-names>
</name>
</person-group> (<year>2013</year>). <article-title>Sin3: insight into its transcription regulatory functions</article-title>. <source>Eur. J. Cell Biol.</source> <volume>92</volume> (<issue>8-9</issue>), <fpage>237</fpage>&#x2013;<lpage>246</lpage>. <pub-id pub-id-type="doi">10.1016/j.ejcb.2013.09.001</pub-id>
</citation>
</ref>
<ref id="B49">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Keogh</surname>
<given-names>M. C.</given-names>
</name>
<name>
<surname>Kurdistani</surname>
<given-names>S. K.</given-names>
</name>
<name>
<surname>Morris</surname>
<given-names>S. A.</given-names>
</name>
<name>
<surname>Ahn</surname>
<given-names>S. H.</given-names>
</name>
<name>
<surname>Podolny</surname>
<given-names>V.</given-names>
</name>
<name>
<surname>Collins</surname>
<given-names>S. R.</given-names>
</name>
<etal/>
</person-group> (<year>2005</year>). <article-title>Cotranscriptional set2 methylation of histone H3 lysine 36 recruits a repressive Rpd3 complex</article-title>. <source>Cell</source> <volume>123</volume> (<issue>4</issue>), <fpage>593</fpage>&#x2013;<lpage>605</lpage>. <pub-id pub-id-type="doi">10.1016/j.cell.2005.10.025</pub-id>
</citation>
</ref>
<ref id="B50">
<citation citation-type="book">
<person-group person-group-type="author">
<name>
<surname>Knight</surname>
<given-names>H. G.</given-names>
</name>
<name>
<surname>Yelon</surname>
<given-names>D.</given-names>
</name>
</person-group> (<year>2016</year>). <source>Utilizing zebrafish to understand second heart field development</source>. <pub-id pub-id-type="doi">10.1007/978-4-431-54628-3_25</pub-id>
</citation>
</ref>
<ref id="B51">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Lagger</surname>
<given-names>G.</given-names>
</name>
<name>
<surname>O&#x27;Carroll</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Rembold</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Khier</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Tischler</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Weitzer</surname>
<given-names>G.</given-names>
</name>
<etal/>
</person-group> (<year>2002</year>). <article-title>Essential function of histone deacetylase 1 in proliferation control and CDK inhibitor repression</article-title>. <source>EMBO J.</source> <volume>21</volume> (<issue>11</issue>), <fpage>2672</fpage>&#x2013;<lpage>2681</lpage>. <pub-id pub-id-type="doi">10.1093/emboj/21.11.2672</pub-id>
</citation>
</ref>
<ref id="B52">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Lazic</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Scott</surname>
<given-names>I. C.</given-names>
</name>
</person-group> (<year>2011</year>). <article-title>Mef2cb regulates late myocardial cell addition from a second heart field-like population of progenitors in zebrafish</article-title>. <source>Dev. Biol.</source> <volume>354</volume> (<issue>1</issue>), <fpage>123</fpage>&#x2013;<lpage>133</lpage>. <pub-id pub-id-type="doi">10.1016/j.ydbio.2011.03.028</pub-id>
</citation>
</ref>
<ref id="B53">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Lei</surname>
<given-names>I.</given-names>
</name>
<name>
<surname>Gao</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Sham</surname>
<given-names>M. H.</given-names>
</name>
<name>
<surname>Wang</surname>
<given-names>Z.</given-names>
</name>
</person-group> (<year>2012</year>). <article-title>SWI/SNF protein component BAF250a regulates cardiac progenitor cell differentiation by modulating chromatin accessibility during second heart field development</article-title>. <source>J. Biol. Chem.</source> <volume>287</volume> (<issue>29</issue>), <fpage>24255</fpage>&#x2013;<lpage>24262</lpage>. <pub-id pub-id-type="doi">10.1074/jbc.M112.365080</pub-id>
</citation>
</ref>
<ref id="B54">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Lei</surname>
<given-names>I.</given-names>
</name>
<name>
<surname>Tian</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Chen</surname>
<given-names>V.</given-names>
</name>
<name>
<surname>Zhao</surname>
<given-names>Y.</given-names>
</name>
<name>
<surname>Wang</surname>
<given-names>Z.</given-names>
</name>
</person-group> (<year>2019</year>). <article-title>SWI/SNF component BAF250a coordinates OCT4 and WNT signaling pathway to control cardiac lineage differentiation</article-title>. <source>Front. Cell Dev. Biol.</source> <volume>7</volume>, <fpage>358</fpage>. <pub-id pub-id-type="doi">10.3389/fcell.2019.00358</pub-id>
</citation>
</ref>
<ref id="B55">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Lickert</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Takeuchi</surname>
<given-names>J. K.</given-names>
</name>
<name>
<surname>Von Both</surname>
<given-names>I.</given-names>
</name>
<name>
<surname>Walls</surname>
<given-names>J. R.</given-names>
</name>
<name>
<surname>McAuliffe</surname>
<given-names>F.</given-names>
</name>
<name>
<surname>Adamson</surname>
<given-names>S. L.</given-names>
</name>
<etal/>
</person-group> (<year>2004</year>). <article-title>Baf60c is essential for function of BAF chromatin remodelling complexes in heart development</article-title>. <source>Nature</source> <volume>432</volume> (<issue>7013</issue>), <fpage>107</fpage>&#x2013;<lpage>112</lpage>. <pub-id pub-id-type="doi">10.1038/nature03071</pub-id>
</citation>
</ref>
<ref id="B56">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Lifschitz</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Haeusler</surname>
<given-names>E. H.</given-names>
</name>
<name>
<surname>Catanho</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Miranda</surname>
<given-names>A. B.</given-names>
</name>
<name>
<surname>Armas</surname>
<given-names>E. M.</given-names>
</name>
<name>
<surname>Heine</surname>
<given-names>A.</given-names>
</name>
<etal/>
</person-group> (<year>2022</year>). <article-title>Bio-strings: A relational database data-type for dealing with large biosequences</article-title>. <source>Biotech. (Basel).</source> <volume>11</volume> (<issue>3</issue>), <fpage>31</fpage>. <pub-id pub-id-type="doi">10.3390/biotech11030031</pub-id>
</citation>
</ref>
<ref id="B57">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Liu</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Yagi</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Saeed</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Bais</surname>
<given-names>A. S.</given-names>
</name>
<name>
<surname>Gabriel</surname>
<given-names>G. C.</given-names>
</name>
<name>
<surname>Chen</surname>
<given-names>Z.</given-names>
</name>
<etal/>
</person-group> (<year>2017</year>). <article-title>The complex genetics of hypoplastic left heart syndrome</article-title>. <source>Nat. Genet.</source> <volume>49</volume> (<issue>7</issue>), <fpage>1152</fpage>&#x2013;<lpage>1159</lpage>. <pub-id pub-id-type="doi">10.1038/ng.3870</pub-id>
</citation>
</ref>
<ref id="B58">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Martinez</surname>
<given-names>S. R.</given-names>
</name>
<name>
<surname>Gay</surname>
<given-names>M. S.</given-names>
</name>
<name>
<surname>Zhang</surname>
<given-names>L.</given-names>
</name>
</person-group> (<year>2015</year>). <article-title>Epigenetic mechanisms in heart development and disease</article-title>. <source>Drug Discov. Today</source> <volume>20</volume> (<issue>7</issue>), <fpage>799</fpage>&#x2013;<lpage>811</lpage>. <pub-id pub-id-type="doi">10.1016/j.drudis.2014.12.018</pub-id>
</citation>
</ref>
<ref id="B59">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>McKinsey</surname>
<given-names>T. A.</given-names>
</name>
</person-group> (<year>2011</year>). <article-title>The biology and therapeutic implications of HDACs in the heart</article-title>. <source>Handb. Exp. Pharmacol.</source> <volume>206</volume>, <fpage>57</fpage>&#x2013;<lpage>78</lpage>. <pub-id pub-id-type="doi">10.1007/978-3-642-21631-2_4</pub-id>
</citation>
</ref>
<ref id="B60">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Monserrat</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Hermida-Prieto</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Fernandez</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Rodriguez</surname>
<given-names>I.</given-names>
</name>
<name>
<surname>Dumont</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Cazon</surname>
<given-names>L.</given-names>
</name>
<etal/>
</person-group> (<year>2007</year>). <article-title>Mutation in the alpha-cardiac actin gene associated with apical hypertrophic cardiomyopathy, left ventricular non-compaction, and septal defects</article-title>. <source>Eur. Heart J.</source> <volume>28</volume> (<issue>16</issue>), <fpage>1953</fpage>&#x2013;<lpage>1961</lpage>. <pub-id pub-id-type="doi">10.1093/eurheartj/ehm239</pub-id>
</citation>
</ref>
<ref id="B61">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Montgomery</surname>
<given-names>R. L.</given-names>
</name>
<name>
<surname>Davis</surname>
<given-names>C. A.</given-names>
</name>
<name>
<surname>Potthoff</surname>
<given-names>M. J.</given-names>
</name>
<name>
<surname>Haberland</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Fielitz</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Qi</surname>
<given-names>X.</given-names>
</name>
<etal/>
</person-group> (<year>2007</year>). <article-title>Histone deacetylases 1 and 2 redundantly regulate cardiac morphogenesis, growth, and contractility</article-title>. <source>Genes Dev.</source> <volume>21</volume> (<issue>14</issue>), <fpage>1790</fpage>&#x2013;<lpage>1802</lpage>. <pub-id pub-id-type="doi">10.1101/gad.1563807</pub-id>
</citation>
</ref>
<ref id="B62">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Morano</surname>
<given-names>I.</given-names>
</name>
<name>
<surname>Chai</surname>
<given-names>G. X.</given-names>
</name>
<name>
<surname>Baltas</surname>
<given-names>L. G.</given-names>
</name>
<name>
<surname>Lamounier-Zepter</surname>
<given-names>V.</given-names>
</name>
<name>
<surname>Lutsch</surname>
<given-names>G.</given-names>
</name>
<name>
<surname>Kott</surname>
<given-names>M.</given-names>
</name>
<etal/>
</person-group> (<year>2000</year>). <article-title>Smooth-muscle contraction without smooth-muscle myosin</article-title>. <source>Nat. Cell Biol.</source> <volume>2</volume> (<issue>6</issue>), <fpage>371</fpage>&#x2013;<lpage>375</lpage>. <pub-id pub-id-type="doi">10.1038/35014065</pub-id>
</citation>
</ref>
<ref id="B63">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Mortensen</surname>
<given-names>S. A.</given-names>
</name>
<name>
<surname>Skov</surname>
<given-names>L. L.</given-names>
</name>
<name>
<surname>Kjaer-Sorensen</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>Hansen</surname>
<given-names>A. G.</given-names>
</name>
<name>
<surname>Hansen</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Dagnaes-Hansen</surname>
<given-names>F.</given-names>
</name>
<etal/>
</person-group> (<year>2017</year>). <article-title>Endogenous natural complement inhibitor regulates cardiac development</article-title>. <source>J. Immunol.</source> <volume>198</volume> (<issue>8</issue>), <fpage>3118</fpage>&#x2013;<lpage>3126</lpage>. <pub-id pub-id-type="doi">10.4049/jimmunol.1601958</pub-id>
</citation>
</ref>
<ref id="B64">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Nakamura</surname>
<given-names>R.</given-names>
</name>
<name>
<surname>Koshiba-Takeuchi</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>Tsuchiya</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Kojima</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Miyazawa</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Ito</surname>
<given-names>K.</given-names>
</name>
<etal/>
</person-group> (<year>2016</year>). <article-title>Expression analysis of Baf60c during heart regeneration in axolotls and neonatal mice</article-title>. <source>Dev. Growth Differ.</source> <volume>58</volume> (<issue>4</issue>), <fpage>367</fpage>&#x2013;<lpage>382</lpage>. <pub-id pub-id-type="doi">10.1111/dgd.12281</pub-id>
</citation>
</ref>
<ref id="B65">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Nambiar</surname>
<given-names>R. M.</given-names>
</name>
<name>
<surname>Ignatius</surname>
<given-names>M. S.</given-names>
</name>
<name>
<surname>Henion</surname>
<given-names>P. D.</given-names>
</name>
</person-group> (<year>2007</year>). <article-title>Zebrafish colgate/hdac1 functions in the non-canonical Wnt pathway during axial extension and in Wnt-independent branchiomotor neuron migration</article-title>. <source>Mech. Dev.</source> <volume>124</volume> (<issue>9-10</issue>), <fpage>682</fpage>&#x2013;<lpage>698</lpage>. <pub-id pub-id-type="doi">10.1016/j.mod.2007.07.003</pub-id>
</citation>
</ref>
<ref id="B66">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Nora</surname>
<given-names>J. J.</given-names>
</name>
</person-group> (<year>1968</year>). <article-title>Multifactorial inheritance hypothesis for the etiology of congenital heart diseases. The genetic-environmental interaction</article-title>. <source>Circulation</source> <volume>38</volume> (<issue>3</issue>), <fpage>604</fpage>&#x2013;<lpage>617</lpage>. <pub-id pub-id-type="doi">10.1161/01.cir.38.3.604</pub-id>
</citation>
</ref>
<ref id="B67">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Papa</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Kushner</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Marx</surname>
<given-names>S. O.</given-names>
</name>
</person-group> (<year>2022</year>). <article-title>Adrenergic regulation of calcium channels in the heart</article-title>. <source>Annu. Rev. Physiol.</source> <volume>84</volume>, <fpage>285</fpage>&#x2013;<lpage>306</lpage>. <pub-id pub-id-type="doi">10.1146/annurev-physiol-060121-041653</pub-id>
</citation>
</ref>
<ref id="B68">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Pitrone</surname>
<given-names>P. G.</given-names>
</name>
<name>
<surname>Schindelin</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Stuyvenberg</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Preibisch</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Weber</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Eliceiri</surname>
<given-names>K. W.</given-names>
</name>
<etal/>
</person-group> (<year>2013</year>). <article-title>OpenSPIM: an open-access light-sheet microscopy platform</article-title>. <source>Nat. Methods</source> <volume>10</volume> (<issue>7</issue>), <fpage>598</fpage>&#x2013;<lpage>599</lpage>. <pub-id pub-id-type="doi">10.1038/nmeth.2507</pub-id>
</citation>
</ref>
<ref id="B69">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Postma</surname>
<given-names>A. V.</given-names>
</name>
<name>
<surname>van Engelen</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>van de Meerakker</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Rahman</surname>
<given-names>T.</given-names>
</name>
<name>
<surname>Probst</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Baars</surname>
<given-names>M. J.</given-names>
</name>
<etal/>
</person-group> (<year>2011</year>). <article-title>Mutations in the sarcomere gene MYH7 in Ebstein anomaly</article-title>. <source>Circ. Cardiovasc Genet.</source> <volume>4</volume> (<issue>1</issue>), <fpage>43</fpage>&#x2013;<lpage>50</lpage>. <pub-id pub-id-type="doi">10.1161/CIRCGENETICS.110.957985</pub-id>
</citation>
</ref>
<ref id="B70">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Pyo</surname>
<given-names>R. T.</given-names>
</name>
<name>
<surname>Sui</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Dhume</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Palomeque</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Blaxall</surname>
<given-names>B. C.</given-names>
</name>
<name>
<surname>Diaz</surname>
<given-names>G.</given-names>
</name>
<etal/>
</person-group> (<year>2006</year>). <article-title>CXCR4 modulates contractility in adult cardiac myocytes</article-title>. <source>J. Mol. Cell Cardiol.</source> <volume>41</volume> (<issue>5</issue>), <fpage>834</fpage>&#x2013;<lpage>844</lpage>. <pub-id pub-id-type="doi">10.1016/j.yjmcc.2006.08.008</pub-id>
</citation>
</ref>
<ref id="B71">
<citation citation-type="book">
<collab>R Core Team</collab> (<year>2021</year>). <source>R: A language and environment for statistical computing</source>. <publisher-loc>Vienna, Austria</publisher-loc>: <publisher-name>R Foundation for Statistical Computing</publisher-name>. <comment>
<ext-link ext-link-type="uri" xlink:href="https://www.R-project.org/">https://www.R-project.org/</ext-link>
</comment>.</citation>
</ref>
<ref id="B72">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Revell</surname>
<given-names>L. J.</given-names>
</name>
<name>
<surname>Graham Reynolds</surname>
<given-names>R.</given-names>
</name>
</person-group> (<year>2012</year>). <article-title>A new Bayesian method for fitting evolutionary models to comparative data with intraspecific variation</article-title>. <source>Evolution</source> <volume>66</volume> (<issue>9</issue>), <fpage>2697</fpage>&#x2013;<lpage>2707</lpage>. <pub-id pub-id-type="doi">10.1111/j.1558-5646.2012.01645.x</pub-id>
</citation>
</ref>
<ref id="B73">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Robinson</surname>
<given-names>M. D.</given-names>
</name>
<name>
<surname>McCarthy</surname>
<given-names>D. J.</given-names>
</name>
<name>
<surname>Smyth</surname>
<given-names>G. K.</given-names>
</name>
</person-group> (<year>2010</year>). <article-title>edgeR: a Bioconductor package for differential expression analysis of digital gene expression data</article-title>. <source>Bioinformatics</source> <volume>26</volume> (<issue>1</issue>), <fpage>139</fpage>&#x2013;<lpage>140</lpage>. <pub-id pub-id-type="doi">10.1093/bioinformatics/btp616</pub-id>
</citation>
</ref>
<ref id="B74">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Severs</surname>
<given-names>N. J.</given-names>
</name>
<name>
<surname>Dupont</surname>
<given-names>E.</given-names>
</name>
<name>
<surname>Coppen</surname>
<given-names>S. R.</given-names>
</name>
<name>
<surname>Halliday</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Inett</surname>
<given-names>E.</given-names>
</name>
<name>
<surname>Baylis</surname>
<given-names>D.</given-names>
</name>
<etal/>
</person-group> (<year>2004</year>). <article-title>Remodelling of gap junctions and connexin expression in heart disease</article-title>. <source>Biochim. Biophys. Acta</source> <volume>1662</volume> (<issue>1-2</issue>), <fpage>138</fpage>&#x2013;<lpage>148</lpage>. <pub-id pub-id-type="doi">10.1016/j.bbamem.2003.10.019</pub-id>
</citation>
</ref>
<ref id="B75">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Shah</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>Seeley</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Schulz</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Fisher</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Gururaja Rao</surname>
<given-names>S.</given-names>
</name>
</person-group> (<year>2022</year>). <article-title>Calcium channels in the heart: disease States and drugs</article-title>. <source>Cells</source> <volume>11</volume> (<issue>6</issue>), <fpage>943</fpage>. <pub-id pub-id-type="doi">10.3390/cells11060943</pub-id>
</citation>
</ref>
<ref id="B76">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Sherman</surname>
<given-names>B. T.</given-names>
</name>
<name>
<surname>Hao</surname>
<given-names>M.</given-names>
</name>
<name>
<surname>Qiu</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Jiao</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Baseler</surname>
<given-names>M. W.</given-names>
</name>
<name>
<surname>Lane</surname>
<given-names>H. C.</given-names>
</name>
<etal/>
</person-group> (<year>2022</year>). <article-title>David: A web server for functional enrichment analysis and functional annotation of gene lists (2021 update)</article-title>. <source>Nucleic Acids Res.</source> <volume>50</volume> (<issue>W1</issue>), <fpage>W216</fpage>&#x2013;<lpage>W221</lpage>. <pub-id pub-id-type="doi">10.1093/nar/gkac194</pub-id>
</citation>
</ref>
<ref id="B77">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Singh</surname>
<given-names>A. P.</given-names>
</name>
<name>
<surname>Archer</surname>
<given-names>T. K.</given-names>
</name>
</person-group> (<year>2014</year>). <article-title>Analysis of the SWI/SNF chromatin-remodeling complex during early heart development and BAF250a repression cardiac gene transcription during P19 cell differentiation</article-title>. <source>Nucleic Acids Res.</source> <volume>42</volume> (<issue>5</issue>), <fpage>2958</fpage>&#x2013;<lpage>2975</lpage>. <pub-id pub-id-type="doi">10.1093/nar/gkt1232</pub-id>
</citation>
</ref>
<ref id="B78">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Song</surname>
<given-names>Y. C.</given-names>
</name>
<name>
<surname>Dohn</surname>
<given-names>T. E.</given-names>
</name>
<name>
<surname>Rydeen</surname>
<given-names>A. B.</given-names>
</name>
<name>
<surname>Nechiporuk</surname>
<given-names>A. V.</given-names>
</name>
<name>
<surname>Waxman</surname>
<given-names>J. S.</given-names>
</name>
</person-group> (<year>2019</year>). <article-title>HDAC1-mediated repression of the retinoic acid-responsive gene ripply3 promotes second heart field development</article-title>. <source>PLoS Genet.</source> <volume>15</volume> (<issue>5</issue>), <fpage>e1008165</fpage>. <pub-id pub-id-type="doi">10.1371/journal.pgen.1008165</pub-id>
</citation>
</ref>
<ref id="B79">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Stankunas</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>Hang</surname>
<given-names>C. T.</given-names>
</name>
<name>
<surname>Tsun</surname>
<given-names>Z. Y.</given-names>
</name>
<name>
<surname>Chen</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Lee</surname>
<given-names>N. V.</given-names>
</name>
<name>
<surname>Wu</surname>
<given-names>J. I.</given-names>
</name>
<etal/>
</person-group> (<year>2008</year>). <article-title>Endocardial Brg1 represses ADAMTS1 to maintain the microenvironment for myocardial morphogenesis</article-title>. <source>Dev. Cell</source> <volume>14</volume> (<issue>2</issue>), <fpage>298</fpage>&#x2013;<lpage>311</lpage>. <pub-id pub-id-type="doi">10.1016/j.devcel.2007.11.018</pub-id>
</citation>
</ref>
<ref id="B80">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Sun</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Hota</surname>
<given-names>S. K.</given-names>
</name>
<name>
<surname>Zhou</surname>
<given-names>Y. Q.</given-names>
</name>
<name>
<surname>Novak</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Miguel-Perez</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Christodoulou</surname>
<given-names>D.</given-names>
</name>
<etal/>
</person-group> (<year>2018</year>). <article-title>Cardiac-enriched BAF chromatin-remodeling complex subunit Baf60c regulates gene expression programs essential for heart development and function</article-title>. <source>Biol. Open</source> <volume>7</volume> (<issue>1</issue>), <fpage>bio029512</fpage>. <pub-id pub-id-type="doi">10.1242/bio.029512</pub-id>
</citation>
</ref>
<ref id="B81">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Takeuchi</surname>
<given-names>J. K.</given-names>
</name>
<name>
<surname>Lou</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Alexander</surname>
<given-names>J. M.</given-names>
</name>
<name>
<surname>Sugizaki</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Delgado-Olguin</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Holloway</surname>
<given-names>A. K.</given-names>
</name>
<etal/>
</person-group> (<year>2011</year>). <article-title>Chromatin remodelling complex dosage modulates transcription factor function in heart development</article-title>. <source>Nat. Commun.</source> <volume>2</volume>, <fpage>187</fpage>. <pub-id pub-id-type="doi">10.1038/ncomms1187</pub-id>
</citation>
</ref>
<ref id="B82">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Trotter</surname>
<given-names>K. W.</given-names>
</name>
<name>
<surname>Archer</surname>
<given-names>T. K.</given-names>
</name>
</person-group> (<year>2008</year>). <article-title>The BRG1 transcriptional coregulator</article-title>. <source>Nucl. Recept Signal</source> <volume>6</volume>, <fpage>e004</fpage>. <pub-id pub-id-type="doi">10.1621/nrs.06004</pub-id>
</citation>
</ref>
<ref id="B83">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>van Engelen</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>Postma</surname>
<given-names>A. V.</given-names>
</name>
<name>
<surname>van de Meerakker</surname>
<given-names>J. B.</given-names>
</name>
<name>
<surname>Roos-Hesselink</surname>
<given-names>J. W.</given-names>
</name>
<name>
<surname>Helderman-van den Enden</surname>
<given-names>A. T.</given-names>
</name>
<name>
<surname>Vliegen</surname>
<given-names>H. W.</given-names>
</name>
<etal/>
</person-group> (<year>2013</year>). <article-title>Ebstein&#x27;s anomaly may be caused by mutations in the sarcomere protein gene MYH7</article-title>. <source>Neth Heart J.</source> <volume>21</volume> (<issue>3</issue>), <fpage>113</fpage>&#x2013;<lpage>117</lpage>. <pub-id pub-id-type="doi">10.1007/s12471-011-0141-1</pub-id>
</citation>
</ref>
<ref id="B84">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>van Oevelen</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Bowman</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Pellegrino</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Asp</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Cheng</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Parisi</surname>
<given-names>F.</given-names>
</name>
<etal/>
</person-group> (<year>2010</year>). <article-title>The mammalian Sin3 proteins are required for muscle development and sarcomere specification</article-title>. <source>Mol. Cell Biol.</source> <volume>30</volume> (<issue>24</issue>), <fpage>5686</fpage>&#x2013;<lpage>5697</lpage>. <pub-id pub-id-type="doi">10.1128/MCB.00975-10</pub-id>
</citation>
</ref>
<ref id="B85">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>van Oevelen</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Wang</surname>
<given-names>J.</given-names>
</name>
<name>
<surname>Asp</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Yan</surname>
<given-names>Q.</given-names>
</name>
<name>
<surname>Kaelin</surname>
<given-names>W. G.</given-names>
<suffix>Jr.</suffix>
</name>
<name>
<surname>Kluger</surname>
<given-names>Y.</given-names>
</name>
<etal/>
</person-group> (<year>2008</year>). <article-title>A role for mammalian Sin3 in permanent gene silencing</article-title>. <source>Mol. Cell</source> <volume>32</volume> (<issue>3</issue>), <fpage>359</fpage>&#x2013;<lpage>370</lpage>. <pub-id pub-id-type="doi">10.1016/j.molcel.2008.10.015</pub-id>
</citation>
</ref>
<ref id="B86">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Voelkel</surname>
<given-names>T.</given-names>
</name>
<name>
<surname>Andresen</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Unger</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Just</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Rottbauer</surname>
<given-names>W.</given-names>
</name>
<name>
<surname>Linke</surname>
<given-names>W. A.</given-names>
</name>
</person-group> (<year>2013</year>). <article-title>Lysine methyltransferase Smyd2 regulates Hsp90-mediated protection of the sarcomeric titin springs and cardiac function</article-title>. <source>Biochim. Biophys. Acta</source> <volume>1833</volume> (<issue>4</issue>), <fpage>812</fpage>&#x2013;<lpage>822</lpage>. <pub-id pub-id-type="doi">10.1016/j.bbamcr.2012.09.012</pub-id>
</citation>
</ref>
<ref id="B87">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Wang</surname>
<given-names>Z.</given-names>
</name>
<name>
<surname>Zhai</surname>
<given-names>W.</given-names>
</name>
<name>
<surname>Richardson</surname>
<given-names>J. A.</given-names>
</name>
<name>
<surname>Olson</surname>
<given-names>E. N.</given-names>
</name>
<name>
<surname>Meneses</surname>
<given-names>J. J.</given-names>
</name>
<name>
<surname>Firpo</surname>
<given-names>M. T.</given-names>
</name>
<etal/>
</person-group> (<year>2004</year>). <article-title>Polybromo protein BAF180 functions in mammalian cardiac chamber maturation</article-title>. <source>Genes Dev.</source> <volume>18</volume> (<issue>24</issue>), <fpage>3106</fpage>&#x2013;<lpage>3116</lpage>. <pub-id pub-id-type="doi">10.1101/gad.1238104</pub-id>
</citation>
</ref>
<ref id="B88">
<citation citation-type="book">
<person-group person-group-type="author">
<name>
<surname>Wickham</surname>
<given-names>H.</given-names>
</name>
</person-group> (<year>2016</year>). <source>ggplot2: Elegant graphics for data analysis</source>. <publisher-loc>New York</publisher-loc>: <publisher-name>Springer-Verlag</publisher-name>.</citation>
</ref>
<ref id="B89">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Xiao</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Gao</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Hou</surname>
<given-names>Y.</given-names>
</name>
<name>
<surname>Xu</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Chang</surname>
<given-names>N.</given-names>
</name>
<name>
<surname>Wang</surname>
<given-names>F.</given-names>
</name>
<etal/>
</person-group> (<year>2016</year>). <article-title>Chromatin-remodelling factor Brg1 regulates myocardial proliferation and regeneration in zebrafish</article-title>. <source>Nat. Commun.</source> <volume>7</volume>, <fpage>13787</fpage>. <pub-id pub-id-type="doi">10.1038/ncomms13787</pub-id>
</citation>
</ref>
<ref id="B90">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Xiao</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Wang</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Hao</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Guo</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Ma</surname>
<given-names>X.</given-names>
</name>
<name>
<surname>Qian</surname>
<given-names>Y.</given-names>
</name>
<etal/>
</person-group> (<year>2018</year>). <article-title>The roles of SMYD4 in epigenetic regulation of cardiac development in zebrafish</article-title>. <source>PLoS Genet.</source> <volume>14</volume> (<issue>8</issue>), <fpage>e1007578</fpage>. <pub-id pub-id-type="doi">10.1371/journal.pgen.1007578</pub-id>
</citation>
</ref>
<ref id="B91">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Yalcin</surname>
<given-names>H. C.</given-names>
</name>
<name>
<surname>Amindari</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Butcher</surname>
<given-names>J. T.</given-names>
</name>
<name>
<surname>Althani</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Yacoub</surname>
<given-names>M.</given-names>
</name>
</person-group> (<year>2017</year>). <article-title>Heart function and hemodynamic analysis for zebrafish embryos</article-title>. <source>Dev. Dyn.</source> <volume>246</volume> (<issue>11</issue>), <fpage>868</fpage>&#x2013;<lpage>880</lpage>. <pub-id pub-id-type="doi">10.1002/dvdy.24497</pub-id>
</citation>
</ref>
<ref id="B92">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Yu</surname>
<given-names>G.</given-names>
</name>
<name>
<surname>Lam</surname>
<given-names>T. T.</given-names>
</name>
<name>
<surname>Zhu</surname>
<given-names>H.</given-names>
</name>
<name>
<surname>Guan</surname>
<given-names>Y.</given-names>
</name>
</person-group> (<year>2018</year>). <article-title>Two methods for mapping and visualizing associated data on phylogeny using ggtree</article-title>. <source>Mol. Biol. Evol.</source> <volume>35</volume> (<issue>12</issue>), <fpage>3041</fpage>&#x2013;<lpage>3043</lpage>. <pub-id pub-id-type="doi">10.1093/molbev/msy194</pub-id>
</citation>
</ref>
<ref id="B93">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Yu</surname>
<given-names>G.</given-names>
</name>
</person-group> (<year>2020</year>). <article-title>Using ggtree to visualize data on tree-like structures</article-title>. <source>Curr. Protoc. Bioinforma.</source> <volume>69</volume> (<issue>1</issue>), <fpage>e96</fpage>. <pub-id pub-id-type="doi">10.1002/cpbi.96</pub-id>
</citation>
</ref>
<ref id="B94">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Zhu</surname>
<given-names>F.</given-names>
</name>
<name>
<surname>Zhu</surname>
<given-names>Q.</given-names>
</name>
<name>
<surname>Ye</surname>
<given-names>D.</given-names>
</name>
<name>
<surname>Zhang</surname>
<given-names>Q.</given-names>
</name>
<name>
<surname>Yang</surname>
<given-names>Y.</given-names>
</name>
<name>
<surname>Guo</surname>
<given-names>X.</given-names>
</name>
<etal/>
</person-group> (<year>2018</year>). <article-title>Sin3a-Tet1 interaction activates gene transcription and is required for embryonic stem cell pluripotency</article-title>. <source>Nucleic Acids Res.</source> <volume>46</volume> (<issue>12</issue>), <fpage>6026</fpage>&#x2013;<lpage>6040</lpage>. <pub-id pub-id-type="doi">10.1093/nar/gky347</pub-id>
</citation>
</ref>
<ref id="B95">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Zhu</surname>
<given-names>L.</given-names>
</name>
<name>
<surname>Vranckx</surname>
<given-names>R.</given-names>
</name>
<name>
<surname>Khau Van Kien</surname>
<given-names>P.</given-names>
</name>
<name>
<surname>Lalande</surname>
<given-names>A.</given-names>
</name>
<name>
<surname>Boisset</surname>
<given-names>N.</given-names>
</name>
<name>
<surname>Mathieu</surname>
<given-names>F.</given-names>
</name>
<etal/>
</person-group> (<year>2006</year>). <article-title>Mutations in myosin heavy chain 11 cause a syndrome associating thoracic aortic aneurysm/aortic dissection and patent ductus arteriosus</article-title>. <source>Nat. Genet.</source> <volume>38</volume> (<issue>3</issue>), <fpage>343</fpage>&#x2013;<lpage>349</lpage>. <pub-id pub-id-type="doi">10.1038/ng1721</pub-id>
</citation>
</ref>
<ref id="B96">
<citation citation-type="journal">
<person-group person-group-type="author">
<name>
<surname>Znosko</surname>
<given-names>W. A.</given-names>
</name>
<name>
<surname>Yu</surname>
<given-names>S.</given-names>
</name>
<name>
<surname>Thomas</surname>
<given-names>K.</given-names>
</name>
<name>
<surname>Molina</surname>
<given-names>G. A.</given-names>
</name>
<name>
<surname>Li</surname>
<given-names>C.</given-names>
</name>
<name>
<surname>Tsang</surname>
<given-names>W.</given-names>
</name>
<etal/>
</person-group> (<year>2010</year>). <article-title>Overlapping functions of Pea3 ETS transcription factors in FGF signaling during zebrafish development</article-title>. <source>Dev. Biol.</source> <volume>342</volume> (<issue>1</issue>), <fpage>11</fpage>&#x2013;<lpage>25</lpage>. <pub-id pub-id-type="doi">10.1016/j.ydbio.2010.03.011</pub-id>
</citation>
</ref>
</ref-list>
</back>
</article>