Fig. 3: Th17 cells generation and gut-homing imprinting of RORγt⁺ pTregs and Th17 cells are impaired by HFD feeding and corrected upon FOS supplementation.

A–C Flow cytometry analysis of total Tregs (A), RORγt⁺ pTregs (B), and Th17 cells (C) in the mesenteric lymph nodes of wild-type mice fed a chow diet (CD) or a high-fat diet (HFD) for 4 weeks (n = 8 mice per group, 2 independent experimental groups). D, E Flow cytometry analysis of RORγt⁺ pTregs (D) and Th17 cells (E) expressing CCR9 or ITGβ7 in the mesenteric lymph nodes of wild-type mice fed a chow diet (CD) or a high-fat diet (HFD) for 4 weeks (n = 7–8 mice per group, 2 independent experimental groups). F, G Flow cytometry analysis of total Tregs (F), RORγt⁺ pTregs (G), and Th17 cells (G) in the mesenteric lymph nodes of wild-type mice fed a high-fat diet (HFD) or a high-fat diet supplemented with FOS (HFD + FOS) for 4 weeks (n = 13–14 mice per group, 3 independent experimental groups). H, I Flow cytometry analysis of CCR9 or ITGβ7-expressing RORγt⁺ pTregs (H) and Th17 cells (I) in the mesenteric lymph nodes of wild-type mice fed a high-fat diet (HFD) or a high-fat diet supplemented with FOS (HFD + FOS) for 4 weeks (n = 13–14 mice per group, 3 independent experimental groups). J, K Flow cytometry analysis of Th17 cells (J) and Th17 cells expressing CCR9 or ITGβ7 (K) in the mesenteric lymph nodes of wild-type mice fed a high-fat diet (HFD) or a high-fat diet supplemented in SCFA (HFD + SCFA) for 4 weeks (n = 4–6 mice per group, 1 experimental group). L, M Flow cytometry analysis of RORγt⁺ pTregs (L) and RORγt⁺ pTregs expressing CCR9 or ITGβ7 (M) in the mesenteric lymph nodes of wild-type mice fed a high-fat diet (HFD) or a high-fat diet supplemented in SCFA (HFD + SCFA) for 4 weeks (n = 4–6 mice per group, 1 experimental group). All data in this figure are presented as mean values ± SEM. Statistical significance has been assessed with a two-sided T test, stars display P value.