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. 2023 Jan 18;55(1):228–239. doi: 10.1038/s12276-022-00913-x

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Comparative RNA-seq analysis shows that 376 genes are differentially expressed (DEG) between mutant (G2-Gata4^Cre/+;Itga4^flox/flox) (n = 4) and control (G2-Gata4^+/+;Itga4^flox/+) (n = 5) E11.5 ventricles (p.value < 1e⁻³); 134 genes are increased and 242 are decreased. Each replicate represents a pool of three ventricles. b Gene Ontology (GO) enrichment analysis (an Over Representation Analysis with a Hypergeometric distribution with a Benjamini–Hochberg adjustment for multiple comparison) reveals important differences between the two genotypes. Pathway enrichment is expressed as the –log [P] adjusted for multiple comparison. The direction of the bars indicates which category was over/underrepresented. All categories in red are overrepresented in mutant animal, whereas all categories in blue are overrepresented in control animals. RT-qPCR analysis of E11.5 embryonic ventricles (n = 5 in control and n = 3 in mutant; each replicate represents a pool of three ventricles) does not reveal significant variations in Vcam1 gene expression (c), but Vcam1 protein mislocalisation is evident in ventricular cardiomyocytes (compare d with e; arrowheads mark the reduced Vcam1 distribution in the lateral plasma membrane of cardiomyocytes). Statistical significance was obtained by a non-parametric one-way analysis of variance with a Kruskal–Wallis post-hoc test (p < 0.05). BrdU uptake (f, g, arrowheads) is significantly reduced in E11.5 mutant ventricular compact myocardium (dashed lines) (n = 5) as compared with stage-matched control (n = 5) (p < 0.01, 3 asterisks, h). The statistical significance was assessed by a t-test and a post hoc Lubischew coefficient analysis. For the quantification of BrdU-positive cells, each replicate represents the mean value from quantifications in three ventricular sections. Abbreviations: BrdU bromodeoxyuridine; CVM compact ventricular myocardium; DAPI diamidino-2-phenylindole; TnI troponin I; TVM trabeculated ventricular myocardium. Scale bars: d, e: 25 µm; f, g: 50 µm.