Figure 4

Reduced CK2 levels in cells treated with gemcitabine result in apoptotic and necrotic cell death. A. Whole lysates from cells treated as indicated in the figure were subjected to Western blot analysis of PARP cleavage. Full length PARP (116 kDa) and the corresponding cleavage product (85 kDa) are indicated. β-actin was used as control for equal loading. B. Cells treated as indicated below the bar graph were subjected to flow cytometry analysis following propidium iodide staining. The amount of cells in sub-G1 is reported in percentage after subtraction of the percentage of cell death in cells treated with transfection reagent. Average from three independent experiments +/- STD is shown. Stars denote statistically significant difference in the percentage of cell death after treatments with CK2α-siRNA and gemcitabine, CK2α'-siRNA or CK2α'-siRNA and gemcitabine as compared to the treatment with gemcitabine alone (Student's t-test, P < 0.03). Statistically significant difference was also found when comparing gemcitabine treatment versus control (results not shown). C. Whole lysates from cells treated as indicated were subjected to Western blot analysis with antibodies directed against a panel of proteins involved in autophagy. For Western blot analysis, three independent experiments were performed obtaining similar results and one representative experiment is shown.