Figure 2.

Gene expression profile and bile acid analysis of male wild type, Fxr^−/−, Tgr5^−/−, and DKO mice, n=6–9. A. QPCR analysis of bile acid synthesis gene expression and FXR-regulated bile acid transporter gene expression in the liver (left) and FXR-induced genes and bile acid transporters in the ileum (right). B. Immunoblotting analysis of liver bile acid synthesis enzyme protein expression. Liver microsomes were isolated to assay CYP7A1 protein and CALNEXIN was used as an internal control. CYP8B1, CYP7B1 and CYP27A1 were assayed in total liver protein and GAPDH was used as an internal control. C. Liver microsomal CYP7A1 enzyme specific activity. D. Total bile acid pool and bile acid concentrations in gallbladder (GB), intestine (Int) and liver (left), serum bile acids (middle) and fecal bile acids (right). E. Conjugated (left) and unconjugated (right) gallbladder bile acid concentrations. WT, wild type mice; Fxr^−/−, FXR single knockout mice; Tgr5^−/−, Tgr5 single knockout mice; DKO, Fxr^−/−/Tgr5^−/− double knockout mice. An “*” indicates statistically significant difference (p<0.05) determined by one-way ANOVA (A, D) or Student’s t-test (B-C, E).