FIGURE 6.

FGF19 inhibits CYP7A1 and other FoxO1 target genes in primary hepatocytes. A, primary mouse hepatocytes were treated with 0, 40 ng/ml, or 80 ng/ml FGF19 for 6 h, and total RNA was isolated and analyzed for expression of the indicted genes by q-PCR as described under “Experimental Procedures.” Data were analyzed as described in the legend to Fig. 4. L32 mRNA levels are shown as a negative control. B, primary mouse hepatocytes were cultured in serum- and glucose-free medium with or without 20, 40, or 80 ng/ml of FGF19 as indicated. After a 3-h incubation, the medium was assayed for glucose production as described under “Experimental Procedures.” p = 0.006, comparing 0 and 80 ng/ml FGF19. C, chromatin was prepared from primary mouse hepatocytes treated with 80 ng/ml FGF19 for 6 h, as described under “Experimental Procedures.” Chromatin immunoprecipitation (ChIP) assays were performed with an anti-FoxO1 or control mouse IgG fraction, and immunoprecipitates were analyzed by q-PCR. Results are expressed as -fold change in comparing the level of DNA amplified from chromatin specifically precipitated by the FoxO1 antibody with that precipitated by a control mouse IgG. The recruitment of FoxO1 to a non-relevant region of the genome in the YY1 locus is shown as a negative control. Data represent the mean of triplicates and include error bars.*, p = 0.02.