Figure 6

Effect of PI3K pathway inhibition on TRG-induced apoptosis resistance in serum-containing media. (A) Huh-7 cells were treated in the absence (-) or presence (+) of 25 μM TRG in serum-containing media for 24 hours following a 1 hour pretreatment with none (lanes 1 & 2), 50 μM LY294002 (lanes 3 & 4), or 20 μM Akt inhibitor VIII (lanes 5 & 6). Western Blot analyses were performed with the antibodies indicated. (B) Cells were treated with TRG as in A following a 1 hour pretreatment with none (lane 2), 50 μM LY294002 (lanes 3), or 20 μM LY303511 (lane 4) and analyzed by Western Blot. (C) Huh-7 cells were treated with TRG as in A, following a 1 hour pretreatment with none (lanes 1 & 2), 10 μM Pak inhibitor (lanes 3 & 4), or 100 nM Rapamycin (lanes 5 & 6). Western Blot analyses with the indicated antibodies were performed next. (D) Huh-7 cells were transfected with either a control-siRNA (lanes 1 & 2), or PPARγ-siRNA (lanes 3 & 4) for 72 hours, followed by TRG treatment in serum-containing media for an additional 24 hours. Western Blot analysis was then performed with the indicated antibodies. TRG-treated Huh-7 cell extract in serum-deficient media was used as positive control for PARP cleavage.