Fig. 2.

A, immunoblotting for total and Ser36 phosphorylated p66^Shc using lysates from PC-3 cells treated with DMSO (control) or 5 μmol/L PEITC for 24 h. B, immunoblotting for total and Ser36 phosphorylated p66^Shc using lysates from PC-3 cells transiently transfected with a control non-specific siRNA or p66^Shc-targeted siRNA and treated for 24 h with DMSO or 5 μmol/L PEITC. The blots were stripped and re-probed with anti-actin antibody to ensure equal protein loading. The numbers on top of the immunoreactive bands represent changes in protein levels relative to corresponding DMSO-treated control (panel A) or relative to DMSO-treated non-specific control siRNA transfected PC-3 cells (panel B). C, cytoplasmic histone-associated apoptotic DNA fragmentation, and D, DCF fluorescence in PC-3 cells transiently transfected with a control non-specific siRNA or p66^Shc-targeted siRNA and treated for 24 h (panel C) or 4 h (panel D) with DMSO or 5 μmol/L PEITC. The results are expressed as enrichment factor relative to DMSO-treated control for PC-3 cells transiently transfected with the control non-specific siRNA. Each experiment was done twice and representative data from a single experiment are shown. Columns, mean (n=3); bars, SE. *Significantly different (P<0.05) between the indicated groups by paired t-test.