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. 2022 Feb 3;145(8):606–619. doi: 10.1161/CIRCULATIONAHA.121.054347

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© 2022 The Authors.

Circulation is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited.

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Figure 6. — ATAC-sequencing shows thousands of newly accessible chromatin overlapping Tbx5 binding sites in Tbx5G125R/+ mice. A, Method of atrial CM nucleus isolation and ATAC-seq. B, Venn diagram showing overlap of accessibility sites in control and Tbx5^G125R/+ mouse atria, as well as the overlap between accessibility sites and Tbx5 binding sites (in %). C, Heatmap clustering showing increasing and decreasing accessibility. Differential accessibility was assessed using the DESeq2 package. P values were corrected for multiple testing using false discovery rate. D, HOMER motif analysis shows reduced accessibility sites in Tbx5^G125R/+ atria were highly enriched for motifs of SP, MEF2, KLF, and TBX families of TFs, whereas Myo family member sites are enriched in increasingly accessible regions. False discovery rate adjusted P values are reported by HOMER. E, ncRNA expression analysis was performed using Limma to determine differential expression. F, University of California, Santa Cruz track showing a ncRNA upstream of Bmp6 (bone morphogenetic protein 6) and differential accessibility. ATAC-seq indicates assay for transposase-accessible chromatin sequencing; CM, cardiomyocyte; DE, differential expression; diff, differential; Eomes (T-box), Eomesodermin; FACS, fluorescence-activated cell sorting; H3K27Ac, histone H3 lysine 27 acetylation; HOMER, Hypergeometric Optimization of Motif Enrichment; Klf10, Krüppel-like factor 10; Klf14, Krüppel-like factor 14; Klf16, Krüppel-like factor 16; Klf3, Krüppel-like factor 3; Klf4, Krüppel-like factor 4; Klf9, Krüppel-like factor 9; Mef2a, myocyte enhancer factor-2a; Mef2b, myocyte enhancer factor-2b; Mef2c, myocyte enhancer factor-2c; Mef2d, myocyte enhancer factor-2d; MyoD, Myogenic Differentiation; MyoG, Myogenin; ncRNA, noncoding RNA; P_adj, P-adjusted value; PCM-1, pericentriolar material-1; SP1, specificity protein 1; SP5, specificity protein 5; Tbet (T-box), T-Box Transcription Factor 21; Tbr (T-box), T-Box Brain Transcription Factor 1; Tbx20, T-box transcription factor 20; TF, transcription factors; and Tn5, transposon 5.