Figure 9.

FoxO1 promotes ATGL promoter activity in an insulin-dependent way. (A) Experimental design outlined. GMECs (8 × 10⁴) were seeded in 48-well plates and incubated overnight. Cells were co-transfected with the ATGL promoter or pRL-TK vector and pcDNA3.1(+) vector or FoxO1-pcDNA3.1, or co-transfected with the ATGL promoter or pRL-TK vector and adenovirus containing FoxO1-specific shRNA or adenovirus containing GFP, and incubation for 12 h. Next, cells were incubated with serum-free medium for 12 h, followed by incubation with or without insulin for 24 h. Finally, GMECs were lysed and used for luciferase activity analysis. Three biological replicates were performed for each experiment and all experiment was individually repeated three times. (B) Relative luciferase activity of ATGL promoter after FoxO1 overexpression in GMECs treatment with or without insulin. GMECs were co-transfected with Ad-GFP or Ad-FoxO1 and ATGL promoter vector for 12 h, followed by 12 h incubation with serum-free medium before being treated with insulin for 24 h. GMECs were lysed and used for luciferase activity analysis. (C) Effects of FoxO1 knockdown on ATGL promoter activity in GMECs treatment with or without insulin. Data are shown as mean ± SEM for three independent replicates. “*”P < 0.05; “NS” P > 0.05.