Fig. 6: VECs contain three main subtypes and Cd36⁺ VECs are markedly increased in hyperlipidemic mice.

a UMAP plot of 536 VECs colored by the clusters and mouse models as indicated. b UMAP plot of VECs color-coded by expression (gray to blue) of marker genes. c Pie chart for the relative proportion of the VEC subsets from each mouse model. d Expression of pro-inflammatory genes in VECs (VEC_C0, C1, and C2). Heatmap (left) and boxplot (right) of the average expression level of genes listed in the heatmap. Heatmap are displayed as expression values scaled by z-transformation on a scale of at least-2.5. Cells having no expression for pro-inflammatory genes were excluded (n = 274 cells for C57BL/6J; 88 for Apoe^−/−; 80 for Ldlr^−/−). Each box depicts the IQR and median of each score, whiskers indicate 1.5 times the IQR. p, two-sided T-test p-value. e Enrichment plot for significant Gene Ontology (GO) terms related to monocyte chemotaxis. Genes were ranked by the fold changes between knockout and wild-type models for cells in VEC clusters (VEC_C0, C1, and C2). f Identification of the localization of VEC subclusters using RNA in situ hybridization (Fgfr3 and Cd36) or immunofluorescence (PROX1). The graph indicates quantification of in situ hybridization using the CD36 probe (n = 5). Two-sided Mann–Whitney test was used. Arrowhead: Cd36⁺ signals in aortic valve. Scale bar: 50 μm. g UMAP plot of VECs color-coded by average expression (gray to red) of genes specific to pre-defined EC subtypes (top). Expression score of genes in EC1 and EC2-associated pathways in VEC_C0 and VEC_C1 (bottom). p, two-sided T-test p-value. h Enrichment map of significant Kyoto Encyclopedia for Genes and Genomes (KEGG) gene sets for each cell cluster. Color represents the adjusted p-value (padj) and size represents the normalized enrichment score (NES), calculated by fgsea R package. Image data are representative of three independent experiments unless otherwise stated. Data are presented as mean ± SD.