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. Author manuscript; available in PMC: 2008 Nov 23.

Published in final edited form as: J Mol Biol. 2007 Sep 18;374(2):334–345. doi: 10.1016/j.jmb.2007.09.031

Cdc2 plays an important role in sensitization to etoposide-induced apoptosis in E6 expressing cells. A, Pharmacologic inhibition of Cdc2 activity reduces apoptosis in E6 expressing cells. NIKS-E6 cells were incubated with either DMSO or etoposide (50 μM) in the presence of Purvalanol A (Calbiochem, 1 μM or 3 μM) for 48 hours. DNA content was determined by flow cytometry after PI staining. The data represent an average of two experiments, each done in duplicate. Error bars represent standard deviations of the means. B, C, and D, Reduction of cdc2 expression by RNAi leads to a decrease of etoposide-induced apoptosis in E6-expressing cells. PHKs expressing E6 were infected with mock adenovirus or adenovirus expressing siRNAs targeting Cdc2 (297) or GFP at a titer of 5 × 10⁶ IFU/ml. Forty-eight hours later, total protein extracts were subjected to Western blot using the Cdc2 antibody (B). In another set of experiments, eight hours after adenovirus infection with GFP siRNA or Cdc2 siRNA, PHKs were treated with etoposide (10 μM) for 48 hours, stained with Annexin V-FITC and PI, and analyzed by flow cytometry. The results of a representative experiment (of 2) showing Annexin V-positive fractions from PI-negative populations are given (C). Dark peaks, DMSO-treated; white peaks, etoposide-treated.

Cdc2 plays an important role in sensitization to etoposide-induced apoptosis in E6 expressing cells. A, Pharmacologic inhibition of Cdc2 activity reduces apoptosis in E6 expressing cells. NIKS-E6 cells were incubated with either DMSO or etoposide (50 μM) in the presence of Purvalanol A (Calbiochem, 1 μM or 3 μM) for 48 hours. DNA content was determined by flow cytometry after PI staining. The data represent an average of two experiments, each done in duplicate. Error bars represent standard deviations of the means. B, C, and D, Reduction of cdc2 expression by RNAi leads to a decrease of etoposide-induced apoptosis in E6-expressing cells. PHKs expressing E6 were infected with mock adenovirus or adenovirus expressing siRNAs targeting Cdc2 (297) or GFP at a titer of 5 × 10⁶ IFU/ml. Forty-eight hours later, total protein extracts were subjected to Western blot using the Cdc2 antibody (B). In another set of experiments, eight hours after adenovirus infection with GFP siRNA or Cdc2 siRNA, PHKs were treated with etoposide (10 μM) for 48 hours, stained with Annexin V-FITC and PI, and analyzed by flow cytometry. The results of a representative experiment (of 2) showing Annexin V-positive fractions from PI-negative populations are given (C). Dark peaks, DMSO-treated; white peaks, etoposide-treated.