Figure 3
Effect of TRG on HCC cell apoptosis in the presence or absence of serum. (A) Subconfluent Huh-7 cells were treated with 25 μM TRG in serum-containing media for the indicated time intervals. Western Blot analysis was performed with antibodies against PARP, Caspase-3 and GAPDH (as control). (B) Huh-7 cells were treated with increasing concentrations of TRG in serum-containing media for 9 hours and subjected to Western Blot analysis as in A. (C) Huh-7 cells were treated with 25 μM TRG in serum deficient media for the indicated time intervals. At the end of incubation cells were harvested and apoptosis assays were performed using cell death detection ELISAPLUS kit. The data in each set represents the mean ± S.D. of 4 independent experiments. (D) Western Blot analysis of cell extracts treated with TRG in serum deficient media for the indicated periods of time and with antibodies against PARP, Caspase-3, cleaved Caspase-3 (detects only the cleaved form) and GAPDH. (E) &(F) Huh-7 cells treated with increasing concentrations of TRG in serum-deficient media were subjected to apoptosis assays using cell death detection ELISAPLUS kit (E) or Western Blot analysis (F).