Figure 4.

FoxO6 regulates hepatic PPAR-γ expression. HepG2 cells were transduced with Adv-FoxO6-CA and control Adv-Empty vectors (200 pfu/cell). After a 24-hour incubation, cells were collected for immunoblot analysis (A). Hepatic protein levels of FoxO6, PPAR-γ, and PGC-1β were determined using actin protein as control (B). Aliquots of cells were used for preparing total RNA, which was subjected to real-time qRT-PCR analysis for determining PPAR-γ mRNA (C) and PGC-1β mRNA levels (D), using β-actin mRNA as control. Furthermore, HepG2 cells were cotransfected with PPREx3-TK-Luc and control pGL4.75 plasmids in the presence of Adv-FoxO6-CA or Adv-Empty control vector. After a 24-hour incubation, cells were subjected to dual luciferase activity assay. PPAR-γ activity was defined as the ratio of Firefly vs Renilla luciferase activity (E). *, P < .05; and **, P < .001 vs control. NS, not significant.