Figure 4.

(A) Antioxidant NAC recovered ROS-dependent apoptosis in NSCLC A549 cells. Cells were evaluated using western blot analysis following pre-treatment with NAC (10 µM) (+) or 0.2% DMSO vehicle control (−) for 1 h, followed by various concentrations of teroxirone treatment for 24 h. The cells were lysed and the protein concentrations determined. Equal amounts of cell lysates and protein were separated by SDS-PAGE and electro-blotted. The blots were subsequently incubated in fresh blocking solution and probed for 1 h with 1:2,000 dilutions of PARP, Akt, p53, Bax, Bcl-2, caspase-3 and GAPDH antibodies, followed by incubation with a 1:3,000 dilution of a horseradish peroxidase-conjugated secondary antibody, prior to being evaluated using an ECL detection system. The numbers below each lane signified relative intensities of cleaved PARP, Akt, p53, Bax, Bcl-2 and GAPDH at each concentration compared with the results of the DMSO vehicle control with or without NAC. (B) NAC recovered ROS-dependent apoptosis in NSCLC H460 cells. Cells were analyzed by western blotting. NAC, N-acetyl cysteine; ROS, reactive oxygen species; NSCLC, non-small cell lung cancer cells; DMSO, dimethyl sulfoxide; PARP, poly ADP-ribose polymerase; Akt, protein kinase B; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2-associated X protein; ECL, enhanced chemiluminescence; p53, tumor protein 53.