Extended Data Fig. 2: Single cell RNAseq analysis and flow cytometric characterization of CD45– PDPN+ CD31– cardiac fibroblasts isolated from 4- and 8-week-old Ctrl and TCRM mice.
(a) Heatmap showing the expression of the top differentially expressed genes from the 10 clusters of cardiac fibroblasts from Ctrl and TCRM mice. (b) Significantly enriched pathways according to Gene Ontology (GO) enrichment analysis based on differentially expressed genes between Ctrl and TCRM cardiac fibroblasts. (c) Flow cytometric gating strategy used to characterize cardiac fibroblast phenotype. (d) Representative density plot depicting the expression of CD157 and NCAM by cardiac fibroblasts from Ctrl and TCRM mice. (e) Differentially regulated functional pathways from single cell RNAseq analysis of cardiac fibroblasts. (f) Expression pattern of genes assigned to the indicated cellular processes projected onto diffusion maps per signature as in panel (e). (g, h) Intracellular phospho-SMAD1/5/9 expression by CD45− CD31− PDPN+ CD56+ cardiac fibroblasts isolated from 4- (N = 4 mice per group) and 8-week-old (N = 5 mice per group) Ctrl and TCRM mice. (g) Representative histogram from cells isolated from 8-week-old mice. (h) Fold change of pSMAD1/5/9 mean fluorescence intensity (MFI) was calculated relative to baseline pSMAD1/5/9 expression in CD56− fibroblasts; pooled data from two independent experiments. Dots indicate individual mice, mean ± SEM is displayed. Statistical analysis was performed using Two-tailed Student’s t test (h). with **, p < 0.01.
