Figure 3. Single-cell Profiling reveals the molecular defects in TGFBR1^A230T CPC-SMC.

A) UMAP projection of the combined dataset of 18,930 scRNA-seq profiles. Individual samples are highlighted in different colors. B) Left: UMAP projection colored based on cluster identity. Right: Fraction of each sample in different clusters (normalized to the total cell number in each sample); TGFBR1^+/+: black, TGFBR1^A230T/+: blue, TGFBR1^A230T/A230T: red. Cell clusters were ranked based on their abundance with C8 being the least abundant cluster in the combined dataset. C) Scaled expression of contractile, and cell proliferation genes on UMAPs. D) Heatmap (scaled by row) showing average expression of binomially specific cluster markers (FDR < 0.01) as well as contractile genes across eight cell clusters. E) Log-transformed FC values for protein-coding genes that are differentially expressed in both TGFBR1^A230T/A230T vs. TGFBR1^+/+ and TGFBR1^A230T/+ vs. TGFBR1^+/+ comparisons (padj < 0.001). F) GSEA showing significantly regulated gene ontology gene sets across two comparisons. Circle sizes indicate the log-transformed p values. NES, normalized enrichment score. G) GSEA showing the cluster-level enrichment or depletion of canonical pathway gene sets from Reactome Pathway Database. The clusters are colored based on log-transformed p-values. H) Percent of EdU+ cells in the control and mutant CPC-SMC. n=5; mean +/− std; *p < 0.05; **p < 0.01; one-way ANOVA with multiple comparison test.