Extended Data Fig. 4: Increased ImP in circulation induces atherosclerosis and systemic inflammation in chow-fed atherosclerosis-prone mice.
From: Imidazole propionate is a driver and therapeutic target in atherosclerosis
a-k, Chow-fed ApoE−/− mice were administered ImP for 4 or 8 weeks as indicated. a, Numbers of RORγt in CD4+ T cells in blood at sacrifice. Left: n = 7. Right: representative density plots for CTR (top) and ImP (bottom). b,c UMAP embeddings of whole aorta-derived cells profiled by scRNAseq colored by cell type, regardless of treatment duration (b) and by treatment and time (c). d, Dot plot of cell type markers used to identify main cell populations. e, UMAP embedding (left) and cell proportions (right) labelled by cell type of whole aorta-derived cells based on transcriptomic profiles from chow-fed ApoE−/− mice after 4 weeks of ImP administration. f, Quantification of atherosclerotic lesions by Oil red O en face staining of the aorta, showing quantification in whole aorta (left), aortic arch (middle) and representative images (right) following 4 weeks of ImP administration. CTR = 10; ImP=8. g-i, Numbers of circulating Ly6Chigh monocytes CTR = 7; ImP=10 (g), CD90+ T cells, B220+ B cells and F4/80+ Macrophages CTR = 9; ImP=10 (h) after 4 weeks of ImP administration determined by flow cytometry. i, Total numbers of CD45+ cells, CD90+ T cells and B220+ B cells infiltrated in aorta after 8 weeks of ImP administration by flow cytometry analysis. CTR = 9; ImP=10. j, Staining of CD3+ cells in intima/media layer in aortic roots showing quantification (left) and representative images (right) of infiltrated T cells after 8 weeks of ImP administration. CTR = 11; ImP=16. Bar size = 500 µm. k, MAC2 staining inside atheroma plaque showing MAC2+ area quantification (left) and representative images (right) after 8 weeks of ImP administration. CTR = 9; ImP=12, Bar size = 500 µm. l, ImP was administered (ImP) or not (CTR) to Ldlr−/− mice grafted with BM from WT (CTR = 9, ImP=10) or Rag1−/− (CTR = 10, ImP=8) and fed chow diet for 12 weeks. Quantification of atherosclerotic lesions by Oil red O en face staining of the aorta, showing quantification in whole aorta. f-l, Individual data and mean ± SEM from at least two pooled independent experiments. a, f (left),h-k, Two-tailed Unpaired Student’s t test. f (right), g, h (left), Two-tailed Mann–Whitney U test. l, One-way ANOVA with Tukey post-hoc correction.
