FIGURE 11.

FZ treatment induces cytochrome c release from mitochondria, activates caspase-3, and causes PARP cleavage. A, H460 cells were treated with either 1 μm FZ for 24 h or 10 μm MG132 for 12 h and then processed for JC-1 staining. a, control; b, FZ; c, MG132. B, after treatment as described in A, the cells were subjected to immunofluorescence using cytochrome c antibody. C, H460 cells were plated onto 35-mm tissue culture plates, and on the following day, the cells were treated with different doses of FZ for 24 or 48 h. The cells were then collected and processed for immunoblotting using caspase-3 and caspase-9 antibodies. D, H460 cells were treated with different doses of FZ or 10 μm MG132 for 48 h, and cell extracts were then prepared and processed for the caspase-3 activity assay using fluorogenic substrate as described under “Experimental Procedures.” E, H460 cell extracts were prepared as described in C, and Western blotting was performed using anti-PARP antibody. Error bars, S.D.