Extended Data Fig. 7: Mice fecal metabolites inhibit Salmonella virulence.
From: Anti-infective bile acids bind and inactivate a Salmonella virulence regulator
a, feces were collected from C57BL/6 mice one day post with or without 20 mg streptomycin treatment. Feces from 4 mice were pooled together and added to lysing matrix D (Methanol: H2O = 9: 1, 100 mg/mL). Feces were then homogenized by Fastprep-24 5 G (MP Biomedicals, 4 m/s, 3 cycles, 5 s for each cycle). Supernatant after centrifugation (18 000 g, 4 oC, 30 min) were lyophilized and the corresponding residues were redissolved in LB for bacterial culture (metabolites from 50 mg of feces were dissolved in 1 mL of LB). Overnight culture of Salmonella (hilA-HA, hilDWT or Q39E+N44D+H95) was subcultured 1:50 to LB with or without metabolites. Bacteria were cultured for 4 h at 37 oC. Expression of HilA-HA and GroEL was analyzed by western blot analysis. b,c, Western blot was quantified by grayscale analysis, normalized with GroEL expression. Bacteria treated with metabolites were compared to bacteria treated with LB by one-way ANOVA and Dunnett’s multiple comparisons test, adjusted P value. Centerline, average. Error bar, SD. (biologically independent samples, n = 3). Comparisons with P > 0.05 were not considered significant.
