Fig. 2.

IL-1β precedes cholestasis and suppresses bile transporters. a Peripheral serum, AST, ALT, total bile acid, and bilirubin concentrations in chow mice, DSS mice immediately after completion of DSS pretreatment (DSS t(0) mice), DSS/chow, DSS/PN, and PN only mice for 3, 7, and 14 days. b Liver mRNA (same mice as in a) of Abcb11 and Abcc2 and in purified intrahepatic mononuclear cells (IHMCs) (c) compared to liver homogenate (d) from chow, DSS t(0), DSS/chow, and DSS/PN mice. e Peripheral serum IL-1β concentrations. f Immunoblot on liver homogenate for procaspase-1 and cleaved caspase-1; protein bands due to non-specific binding of this antibody and for anti-GRB2 antibody were used to show equal protein loading. For the latter, the blot needed to be stripped due to proximity in size of GRB2 (25 kDa) and IL-1β (17 kDa active, 31 kDa pro-form). g Liver mRNA of Abcb11 and Abcc2 from untreated and IL-1β-injected mice after 4 h in Abcb11 mRNA and Abcc2 mRNA in HuH7 cells (h) or primary mouse hepatocytes (i) left untreated or exposed to IL-1β (10 ng/ml) for 4 h. For h and i representative data are shown depicting mean ± SEM of technical triplicates from one out of at least five experiments. j, k Il1b, Abcb11, and Abcc2 mRNA expression in liver homogenate or in IHMCs from untreated and LPS injected mice after 4 h with or without prior (16 h) i.p. clodronate-liposome (Clo-lip) injection and in WT, Casp1/11^−/−, orIl1r^−/− mice (l). *p < 0.05 vs. all other groups by one-way ANOVA and Tukey's correction (a, b, e). *p < 0.05 vs. chow, and ^#p < 0.05 vs. chow/DSS and chow (c, d). *p < 0/05 vs. untreated by t test (g, h, i, j); *p < 0.05 vs. untreated and LPS-Clo-lip one-way ANOVA and Tukey's correction (k); *p < 0.05 vs. untreated, and ^#p < 0.05 vs. WT by one-way ANOVA and Tukey's correction (l). Data are depicted as individual mice for serum analysis and as means from PCR triplicates from individual mice for gene expression data (mean ± SEM)