Fig. 4: EndMA is a reversible activation of endothelial cells.

a tSNE plot showing all sequenced cells for all timepoints (Homeostasis, d1, d3, d7, d14, d28) in hearts of tamoxifen treated Cdh5-CreERT2;mT/mG animals (n = 15,365 cells, 1 mouse per time point). We found four mesenchymal clusters (0,1,3,9), three endothelial clusters (2,8,13), three macrophage/monocyte clusters (4,7,11), four clusters of smooth muscle cells/pericytes (10,14,15,16), two clusters of T-cells (6,12), and one cluster of B-cells (5). b tSNE plot showing GFP⁺ traced endothelial cells (n = 1393 cells). c tSNE plot highlighting expression levels of endothelial marker (Pecam1, Cdh5, Vwf) and mesenchymal marker (Col1a1, Pdgfra, Serpine1) as scaled normalized UMI. d Left panel (green) displays relative number of cells expressing (UMI ≥ 1) GFP, Cdh5, and the mesenchymal markers (Col1a1, Serpine1, or Col3a1) per timepoint. Right panel (blue) shows population of cells not expressing Cdh5 in this context. Data represents n = 1 mouse per timepoint. Homeostasis n = 592 cells, d1 n = 87 cells, d3 n = 44 cells, d7 n = 166 cells, d14 n = 304 cells, d28 n = 270 cells. e tSNE plot of reclustered GFP positive cells, showing 7 independent clusters. f Gene expression of different marker genes in different cluster shown in e. Color indicates the log-fold change (blue to red), size shows log p-value. g Relative number of cells assigned to the different clusters from (e) in homeostasis and d7. h, i Fold change of SM22 (TAGLN) (h) and Calponin (CNN1) (i) mRNA levels measured by qRTPCR (n = 3 independent experiments) in HUVECs treated with TGF-β2 supplemented medium (turquoise) and control medium (gray). Expression values were normalized to RPLP0. HUVECs cultured for 3 days in TGF-β2 and subsequent cultivation in control medium until 7 days are shown in green. P-value was calculated using Kruskal–Wallis test (p = 0.005 SM22, p = 0.006 CNN1). Data shown as mean ± SEM.