Figure 3.

FoxO3a overexpression increases tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) expression. Cells were stably transfected with a control vector (pLenti/V5-D-TOPO) or cDNA encoding wild-type FoxO3a. (A) FoxO3a expression was assessed by Western blotting using anti-FoxO3a Ab. (B) Control vector or FoxO3a overexpressing cells were changed to serum-free medium. After 9 h, cells were incubated with synthetic oligodeoxynucleotides (ODN) with the CpG-motif (CpG ODN) for 6 h, and TRAIL mRNA levels were assessed by real-time PCR. (C) Vector or FoxO3a overexpressing cells were changed to serum free media and treated with 3 μM CpG ODN for 12 h, stained with propidium iodide (PI), and analyzed for the induction of apoptosis by flow cytometry. Each data point represents the mean ± SD of three independent experiments (^*P<0.05 and ^**P<0.01).