Figure 3.

Shared and cell type–specific differentially expressed genes (DEGs) induced by trimethylamine-N-oxide (TMAO) in major aortic cell types. A, Shared and cell type–specific DEGs (false discovery rate [FDR] <0.05) induced by TMAO feeding in major aortic cell types recovered in single-cell RNA-sequencing. DEGs unique to a cell type are highlighted in red in the upset plot and histogram, and shared DEGs are indicated in black. The histogram above each plot indicates the DEG counts for each category. DEG direction is indicated by the color of the gene name: red: upregulated in high-cholesterol (HC)+TMAO vs HC, blue: downregulated in HC+TMAO vs HC, black: cell type–dependent. B and C, Top representative pathways from Reactome enriched for upregulated and downregulated modulated vascular smooth muscle cell (vSMC) DEGs (FDR<0.05), respectively. D, ATP8 and ND3 expression in immortalized human aortic smooth muscle cells with TMAO stimulus. E, COL1A1, COL1A2, TGFB1, and TP53INP1 gene expression in primary human coronary artery smooth muscle cells with TMAO stimulus. One-way ANOVA with Tukey post hoc test was used to determine statistical significance.