Fig. 5: Endothelial Rnf20 loss results in defects in EC differentiation and ventricular CM proliferation. | Nature Communications

Fig. 5: Endothelial Rnf20 loss results in defects in EC differentiation and ventricular CM proliferation.

From: Endothelial RNF20 suppresses endothelial-to-mesenchymal transition and safeguards physiological angiocrine signaling to prevent congenital heart disease

Fig. 5

a Schematic representation of the heart dissection procedure used for scRNA-Seq experiments. Illustration of the E11.5 embryo created in BioRender.com. Dobreva, G. (2025) https://BioRender.com/56nm0tt. Hearts from five control and Rnf20iEC-KO embryos (n = 5) were dissected for the experiment. b t-SNE plot of 10x genomics scRNA-Seq datasets of E11.5 control and Rnf20iEC-KO hearts. OFT/AVCm1 and 2, outflow tract/atrioventricular canal mesenchyme 1 and 2; dOFTm, dorsal outflow tract mesenchyme; Ery, erythrocytes; EC/EndoC, endothelial cells/endocardial endothelial cells; CM1 and CM2, cardiomyocytes 1 and 2; pMES, proliferative mesenchyme; EpiC1 and EpiC2, epicardial cells 1 and 2; OFTe, outflow tract epithelium; EndoV, endocardial valve cells; HSPC, hematopoietic stem and progenitor cells. c Percentage of cells within the different clusters colored by cell types. P-values were calculated using a two-sided exact binomial test. d dot plot depiction of representative gene ontology terms in up- and downregulated genes in the different CM and EC populations of Rnf20iEC-KO versus control hearts. e UMAP re-clustering of CM1 and CM2 from panel b, colored by CM type. vCM1 and vCM2, ventricular cardiomyocytes 1 and 2; pvCM1-2, proliferative ventricular CM1-2; mvCM mature ventricular CM, aCM1 and aCM2, atrial cardiomyocytes 1 and 2, aCMim immature atrial cardiomyocytes, AVC/OFT atrioventricular canal/outflow tract, OFT outflow tract. f Feature plots of Mki67 expression in the different CM populations in (e). Values represent normalized gene expression counts per cell. g Percentage of cells in each CM subtype, color-coded by cell type. P-values were calculated using a two-sided exact binomial test. h UMAP analysis of Pecam1+ECs, showing five distinct EC clusters. i Bar plot showing the percentage of cells across different EC types. P-values were calculated using a two-sided exact binomial test. j Dot plot displaying EC and EndMT marker gene expression in the EC clusters. EndoC endocardial ECs, pEndoC proliferative endocardial ECs, EndoV1-2 endocardial valve cells 1 and 2, VECs vascular ECs. k Representative GO terms in up- and downregulated genes in Rnf20iEC-KO in the different EC populations in (h).

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