Fig. 3: Increased circulating ImP induces atherosclerosis and systemic inflammation in chow-fed atherosclerosis-prone mice.
From: Imidazole propionate is a driver and therapeutic target in atherosclerosis
a, ImP was administered (ImP) or not (Ctrl) to chow-fed Ldlr−/− mice. Oil red O staining of aortic arch (left, n = 10) and representative images of aorta (right). b–e, ImP was administered (ImP) or not (Ctrl) to Apoe−/− mice. b, Quantification (left) and representative images (right) of Oil red O staining of aortic root lesions. n = 12. Arrowheads indicate areas with positive Oil red O staining. Scale bars, 500 µm. c,d, Flow cytometry analysis of Ly6Chi monocytes (n = 8) (c) and T-bet in CD4+ T cells (n = 6) (d) in blood at endpoint. Right, representative density plots for Ctrl and ImP. e, Uniform manifold approximation and projection (UMAP) embedding (left) and cell proportions (right) labelled by cell type of cells derived from whole aorta, based on transcriptomic profiles. DCs, dendritic cells; ECs, endothelial cells; FBs, fibroblasts; MFs, macrophages; VSMCs, vascular smooth muscle cells. f, Ldlr−/− mice grafted with bone marrow from wild-type (WT) (Ctrl, n = 9; ImP, n = 10) or Rag1−/− (Ctrl, n = 10; ImP, n = 8). Atherosclerosis by Oil red O staining of aortic arch (left) and representative images of aorta (right). g, Heat map of GSEA comparing ImP versus control for endothelial cells, fibroblasts and macrophages from scRNA-seq data of Apoe−/− mice fed chow diet for 4 weeks. EMT, epithelial–mesenchymal transition; NES, normalized enrichment score. h, Global abundance of peptides (mTOR) and phosphopeptides (p-mTOR) in BMDMs (left) or MEFs (right) treated with ImP for 3 h with or without AGN192403 (AGN). The log2 ratio is relative to unstimulated conditions. i, Ldlr−/− mice grafted with bone marrow from control Raptorfl/fl (Ctrl, n = 17; ImP, n = 18) or Lyz2ΔRaptor (Ctrl, n = 15; ImP, n = 18). Quantification (left) and representative images (i) of Oil red O staining of aorta, showing atherosclerosis in aortic arch. a–d,f,i, Individual data points and mean ± s.e.m. of at least two pooled independent experiments. a–d, Two-tailed unpaired Student’s t-test. f,i, One-way ANOVA with Tukey post hoc correction. g, Two-tailed Kolmogorov–Smirnov test comparing ImP versus unstimulated. h, Two-tailed Wilcoxon signed rank test comparing peptide and phosphopeptide abundance in ImP versus ImP plus AGN treatments.
