Figure 1.

Identification of major aortic cell types and cell type–specific gene markers. A, Schematic diagram of the study design. Athero-prone areas of the aorta (ascending aorta, aortic arch, and thoracic aorta) were collected from female Ldlr^−/− mice on normal laboratory (NL), high-cholesterol (HC), or HC+ trimethylamine-N-oxide (TMAO) diets (n=6/group), and 2 mouse samples from the same diet group were pooled together, with n=3 pools/group. B, Plasma TMAO levels (µM) in the NL-, HC-, and HC+TMAO-fed mice as measured by mass spectrometry (n=6/group). Statistical significance was determined by unpaired t test. C through E, Uniform Manifold Approximation and Projection (UMAP) representation of cell clusters in NL, HC, and HC+TMAO conditions, respectively. F, Cluster-specific expression of previously known cell type markers. G, Normalized expression values of top markers of vascular smooth muscle cells (vSMC) subtype clusters: vSMC 1: Acta2, vSMC 2: Atf3, Rgs5⁺ vSMC: Rgs5, modulated vSMC: Spp1. H, Proportions of identified cell types within total cells recovered for each diet condition in order of abundance. Statistical significance was determined by unpaired t test. False discovery rate (FDR) was calculated with Benjamini-Hochberg. DEG indicates differentially expressed genes; and Mod. vSMC, modulated vSMC.