Enriched vascular gene profile emerges in cripDM embryos. (A) Dot plot representation for the cell type identification of endothelial populations using key arterial (dll4, hey2, flt1, notch1b, efnb2a), venous (flt4, mrc1a, dab2, stab2) and cell cycle (kif23, mki67, nusap1, tacc3) marker genes. Cell types include endothelial venous (EC venous), arterial-venous (AV), endothelial arterial (EC arterial), endothelial venous proliferating and endothelial arterial proliferating. Expression bar illustrates scaling performed per gene where the mean is close to 0 and the standard deviation is 1. Only genes with the highest scaled expression value will be dark purple (s.d.>1) and the lowest scaled expression will be light purple (s.d.<−1). Percent of cells expressing gene in the population is depicted by the size of the dot. (B) Cell type classifications of ECs in UMAP space as identified in A. (C) Volcano plot for upregulated (red, right) and downregulated (blue, left) genes between wild-type and cripDM embryos in total EC population. Horizontal red line is at a -log10(P-value adjusted) value of 0.005 and vertical red lines are at a mean log2(fold change) value of −0.15 and 0.15. (D) Violin plots for the vasculature development gene expression score for the endothelial populations in wild-type and cripDM embryos. Pairwise Wilcox test (P<0.0001). (E) Gene Ontology enrichment scores between wild-type and cripDM samples for arterial ECs. (F-K) In situ hybridization for dll4 in wild-type (n=41) (F) and cripDM (n=41) (G) embryos and for notch1b in wild-type (n=38) (I) and cripDM (n=40) (J) embryos at 30 hpf. Lateral views, anterior to the left. Scale bars: 50 µm. Violin plots for dll4 (H) and notch1b (K) gene expression in scRNA-seq dataset for endothelial populations in wild-type and cripDM embryos. Pairwise Wilcox test was used for these analyses (H,K) and yields statistically significant differences in noted venous (****P<0.0001), arterial-venous (****P<0.0001) and arterial (*P<0.05) populations. ns, not significant.
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