Fig. 5: Itaconate production sustains gut microbiota alterations underlying HFD-induced obesity.

A Fecal itaconate levels of WT and Acod1^-/- mice fed for 16 weeks with ND or HFD. Data as mean±s.e.m. of n = 12 mice per group from three experiments independently executed. B Fractional analysis of the fecal bacterial composition at the phylum level of mice described in A. Data as mean±s.e.m. of n = 10 (ND) or n = 15 (HFD) mice from three independent experiments. Differential abundance analysis (DESeq2) of the fecal bacterial composition at family (C) and genus (D) level of WT (n = 15) and Acod1^-/- (n = 15) mice fed with HFD for 16 weeks from three independent experiments. Dashed line indicates FDR = 0.05. The size of the dots indicates fold change (log2) and the color indicates the phylum. E Levels of fecal SCFAs in mice treated as in A. Data as mean±s.e.m. of n = 7 (ND-fed WT), n = 6 (ND-fed Acod1^-/-), n = 8 (HFD-fed WT), n = 11 (HFD-fed Acod1^-/-) mice from two experiments independently executed. F Fractional analysis of the fecal bacterial composition at the phylum level of HFD-fed WT mice treated with 20 mM itaconate in drinking water for 12 weeks. Data as mean±s.e.m. of n = 10 mice per group from two experiments independently executed. Differential abundance analysis (DESeq2) comparing fecal bacterial composition at family (G) and genus (H) levels of mice described in F (n = 10 mice per group from two experiments independently executed). Dashed line indicates FDR = 0.05. The size of the dots indicates fold change (log2) and the color indicates the phylum. I Effect of itaconate (0.5 mM) treatment on growth (OD 600 nm) of five pure bacteria strains representative of the Bacteroidaceae family (Bacteroides genus), over 24 h of culture in anaerobic atmosphere, measured spectrophotometrically. Data as mean ± s.e.m. of n = 6 samples per condition from two independent experiments. J Body weight of HFD-fed WT mice treated with 20 mM itaconate in drinking water for the time indicated. Data as mean ± s.e.m. of n = 10 (vehicle) and n = 12 (itaconate)-treated mice from two experiments independently executed. K Tissue weights from mice escribed in J. Data as mean±s.e.m. of n = 10 (vehicle) and n = 12 (itaconate)-treated mice from two experiments independently executed. L ITT in WT mice treated with 20 mM itaconate in drinking water for 12 weeks. Data as mean±s.e.m. of n = 5 (vehicle) and n = 7 (itaconate)-treated mice from one representative experiment out of two independently executed. In: (A), (B), (E), (F), (I) and (K) t-test. In (J) and (L) two-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001; ns not significant.