FIG. 7.

Zyflamend induced up-regulation of DR5 and CHOP is mediated through generation of ROS. (A-B) HCT-116 cells were treated with indicated concentrations of NAC (A) or GSH (B) for 1 h, and then treated with 250 μg/mL Zyflamend for 24 h. Whole-cell extracts were prepared and analyzed by Western blotting using indicated antibodies. β-Actin was used as a loading control. (C-D) Zyflamend-modulated anti-apoptotic and pro-apoptotic proteins expression is dependent on ROS. HCT-116 cells were pre-treated with NAC or GSH for 1 h and then treated with Zyflamend (250 μg/mL) for 24 h. Whole-cell extracts were prepared and analyzed by Western blotting using the relevant antibodies. β-Actin was used as an internal control to verify equal protein loading. (E) Zyflamend-induced DR5 and CHOP expression in pancreatic cancer cells and human embryonic kidney cells is mediated through ROS. Cells were first treated with NAC or GSH for 1 h and then with 250 μg/mL Zyflamend for 24 h. Whole-cell extracts were prepared and analyzed by Western blotting using indicated antibodies. β-Actin was used as a loading control.