Figure 3

The effect of P276-00, gemcitabine and the combination of gemcitabine followed by P276-00 on (A). Cells were treated with 300 nM P276-00 for 72 h or 70 nM of gemcitabine for 24 h followed by only medium for additional 72 h or combination treatment of gemcitabine for 24 h followed by P276-00 for 72 h. (B) Cells were treated with 300 nM P276-00 for 96 h or 70 nM of gemcitabine for 24 h followed by only medium for additional 96 h or combination treatment of gemcitabine for 24 h followed by P276-00 for 96 h. Cell cycle analysis using Flow cytometry shows the percentage apoptosis in drug alone and combination treatment in comparison to untreated control. The percent distribution of cells in various phases of the cell cycle is represented as bar plot (C) Western blot analysis in PANC-1 cells of the antiapoptotic proteins Bcl-2 and survivin. Samples were obtained from PANC-1 cells treated with 300 nM of P276-00 for 72 h or gemcitabine for 24 h or the combination of gemcitabine for 24 h followed by P276-00 for 72 h. (D) Densitometric analysis of the Bcl-2 and survivin bands using the software Image J.