Fig. 2: Fructooligosaccharides supplementation improves glucose tolerance and prevents RORγt⁺ pTregs and Th17 cells loss in HFD-fed animals.

A–E Body weight (n = 15 mice per group) (A), body weight gain (n = 15 mice per group) (B), epididymal fat mass (n = 14–15 mice per group) (C), and body composition (fat and lean mass) (n = 7–10 mice per group. Statistical significance assessed with 2-way ANOVA and Sidak’s multiple comparison test, stars display adjusted P value) (D) and daily food intake (n = 9 mice per group) (E) in wild-type mice fed a high-fat diet (HFD) or a high-fat diet supplemented with fructooligosaccharides (FOS) (HFD + FOS) administered in the drinking water for 4 weeks. F–J Oral glucose tolerance test and associated area under the curve (AUC) quantification (n = 15 mice per group) (F), HOMA-IR index measurement (n = 15 mice per group) (G), plasma insulin levels (n = 15 mice per group) (H), fasted plasma glucose levels (n = 15 mice per group) (I) and glucose-stimulated insulin secretion (GSIS) (n = 9–12 mice per group, 2 independent experimental groups) (J) in wild-type mice fed a high-fat diet (HFD) or a high-fat diet supplemented with FOS (HFD + FOS) for 4 weeks. K–M Colon weight (n = 13–14 mice per group) (K), colon length (n = 9–12 mice per group) (L), and cecum weight (n = 13–14 mice per group, two independent experimental groups) (M) in wild-type mice fed a high-fat diet (HFD) or a high-fat diet supplemented with FOS (HFD + FOS) for 4 weeks. N–P Flow cytometry analysis of total Tregs (N), RORγt⁺ pTregs (O), and Th17 cells (P) in the small intestine and colon 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). Q–X Microbiome sequencing and analysis of phyla relative abundance (Q), Shannon index of microbial diversity (R), relative abundance of specific bacterial class or species including Segmented Filamentous Bacteria (SFB) (S), Bifidobacteria (T), Clostridia (U), Fusobacterium nucleatum (V), Bifidobacterium breve (W), and Staphylococcus saprophyticus (X) in wild-type mice fed a chow diet (CD), a high-fat diet (HFD) or a high-fat diet supplemented with FOS (HFD + FOS) for 4 weeks (n = 4 mice per group, each from an independent experimental group. Statistical testing with one-way ANOVA and Newman-Keuls multiple comparison test). All data in this figure are presented as mean values ± SEM. All panels correspond to three independent experimental groups unless otherwise specified in the corresponding panel legend. Statistical significance has been assessed with a two-sided T test unless otherwise stated in the corresponding panel legend.