FIGURE 8.

Insulin-suppressed expression of key autophagy genes was mediated by FoxO1. The constitutive nuclear form of FoxO1 (ADA-FoxO1) encoded by the recombinant adenovirus or the empty adenoviruses was introduced into Hepa1c1c7 cells via infection. Thirty-six hours later, cells were cultured in the starvation medium (EBSS) in the presence or absence of insulin (1 nm) for an additional 8h. A, levels of LC3, phosphorylated Akt, and β-actin were measured by immunoblotting. Levels of FoxO1 and PARP1 in nuclear protein extracts were measured by immunoblotting with the antibodies as noted. *, p < 0.05. B, transcripts of vps34, atg12, and gabarapl1 genes were determined by using real-time PCR and normalized to the level of 36B4 transcript. Results represent mean ± S.E. from three independent experiments. *, p < 0.05; **, p < 0.01 insulin plus starvation versus starvation only. #, p < 0.05; ###, p < 0.001 ADA-FoxO1 infection plus insulin plus starvation group versus starvation only group.