Extended Data Fig. 5: SI T_RM uptake and sense dietary cholesterol.

a, Quantification by flow cytometry of in vivo incorporation of Bodipy-cholesterol on indicated cell populations after 10 minutes after oral administration of Bodipy-cholesterol to memory P14 mice. LPL, lamina propia lymphocytes. b, Quantification by flow cytometry of incorporation of Bodipy-LDL on indicated cell types of the spleen 20 minutes (min) after IV delivery to memory P14 mice. c, Quantification of intracellular cholesterol by filipin staining in Tet⁺ populations of CD8⁺ T cells for various tissues >30 days post LCMV infection measured by imaging flow cytometry. d, Gene expression analysis by qPCR of selected SREBP2 targets on CD8⁺ T cells from the spleen and SI of mice on regular chow or subjected to low and high cholesterol-containing diets. e, Experimental design to measure transcriptional changes induced by dietary cholesterol on established SI T_RM by Ultralow-input RNA-seq, f, SI T_RM frequencies from total CD8⁺ T cells after 7 days of dietary intervention. g, Food consumption per day per mouse subjected to a low or high-cholesterol diet. h, Bodyweight of mice subjected to low or high cholesterol-containing diets. i, Total cholesterol in serum in mice subjected to low or high cholesterol-containing diets. j, Volcano plot of differentially expressed genes in SI T_RM from mice fed a high vs a low cholesterol-containing diet. k, Mevalonate-cholesterol synthesis GSVA scores in SI T_RM from mice fed a high versus low cholesterol-containing diet. l, Gene pathway analysis of top upregulated and top downregulated genes between SI T_RM from mice fed a high versus low cholesterol-containing diet (MsigDb, Hallmark gene sets). P14 CD8⁺ T cells and CD8β⁺/CD44^high/Tet⁺ from the kidney, liver and SI were gated on the IV⁻ population (a, b, c, e, f, j, k, and l). Data are mean +/− s.e.m. and representative of at least two independent experiments, with a total of n = 4 (a), n = 3 (b, d), n = 5 (c), n = 3 (f) mice, or pooled from at least two independent experiments with a total of n = 5–10 (g-i) mice, or performed once (j-l) with a total of n = 3 samples each sample is 1 × 10³ SI T_RM pooled from 2~3 mice (j, k). Two-sided unpaired t-Test (a, b, d, f, i, and k), and Deseq2 DEG testing with Benjamini-Hochberg multiple test correction (j). *P < 0.05, **P < 0.01, ***P <  0.005. The images of the mice in a,b,e have been modified from ref. ¹⁷.

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