Figure 6.

USP7's effect on gluconeogenic gene expression is dependent on FoxO1. A, Efficient knockdown of FoxO1 protein upon double knockdown with USP7. Western blot analysis of whole-cell extracts from primary hepatocytes infected with the indicated adenoviruses (in the absence of shFoxO1, cells received coinfection with corresponding control shRNA adenovirus) and serum starved in 0.2% BSA for 3 hours prior to a 1.5 hours incubation with 10 μM forskolin. B, Knockdown of FoxO1 abolishes the effect of USP7 knockdown on gluconeogenic gene expression. mRNA levels are graphed relative to expression of unstimulated/serum-starved control shSCR and presented as average ± SEM; n = 3. *, P < .05; and ***, P < .001 by one-way ANOVA with Newman-Keuls Multiple Comparison test. Data are representative of at least two independent experiments.