FIG. 8.

Potentiation of TRAIL-induced apoptosis by Zyflamend is ROS dependent. (A-E) NAC and GSH reversed cell death induced by the combination of Zyflamend and TRAIL in cancer cells. Cells were pre-treated with NAC or GSH for 1 h before treatment with Zyflamend. After 12 h, cells were washed with PBS and then treated with TRAIL for 24 h. Cell death was determined by the live/dead assay. (F) Whole-cell extracts prepared from treated cells were analyzed by Western blotting using antibodies against caspase-8, caspase-9, and caspase-3. β-Actin was used as an internal control to verify equal protein loading. (G) Zyflamend induces expression of DR5, CHOP and down-regulates anti-apoptotic proteins in vivo. The protein extract obtained from the tumor tissues of vehicle treated and Zyflamend treated nude mice were analyzed by Western blot analysis using indicated antibodies. (To see this illustration in color the reader is referred to the web version of this article at www.liebertonline.com/ars).