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. 2010 Aug 26;42(10):712–720. doi: 10.3858/emm.2010.42.10.070

Figure 8.

Figure 8

LY294002 is inhibited CpG ODN-mediated anti-apoptosis through FLIP expression. (A) Cells were serum starved for 12 h, incubated with 3 µM CpG ODN for the indicated times, and Akt phosphorylation was assessed by Western blotting using Akt and phospho Akt Ab. (B) Cells were incubated with 3 µM CpG ODN for 30 min in the absence or presence of 10 µM LY294002. FoxO3a phosphorylation was then assessed by Western blotting using FoxO3a and phospho FoxO3a Ab. (C) Cells were serum starved for 12 h, pretreated with 10 µM LY294002, and incubated with 3 µM CpG ODN. FLIP mRNA levels were assessed by real-time RT-PCR. (D) Cells were changed into serum-free medium, pretreated with 10 µM LY294002, and incubated with 3 µM CpG ODN for 12 h. Cells were stained with annexin-V FITC, and analyzed for the induction of apoptosis by flow cytometry. Each data point represents the mean ± S.D. of four independent experiments (*, P < 0.05).