a, b, IsoLCA inhibited the differentiation of mouse TH17 cells in vitro. Representative FACS plots (a) and population frequencies of TH17 cells (b) cultured in the presence of various BAs (20 μM) and ML209 (2 nM) (n = 3 biologically independent samples per condition, data are mean ± SEM, one-way ANOVA followed by Dunnett’s multiple comparison test, vehicle set as control).
c-f, IsoLCA inhibited the differentiation of TH17 cells in vivo. Experimental scheme (c), representative FACS plots (d) and population frequencies of TH17 cells (e) and Treg cells (f) in the ileal lamina propria of SFB-colonized mice are shown. B6 Jax mice were gavaged with SFB-rich fecal pellets and maintained on control or isoLCA-containing powder chow (0.3% w/w) for one week (n=8 mice per group, pooled from two experiments, data are mean ± SEM, two-tailed unpaired t-test).
g, Three-dimensional structures of 3-oxoLCA, isoLCA, and LCA showing the facial orientation of the C3 oxygenation.