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. 2011 Aug 24;6(8):e23849. doi: 10.1371/journal.pone.0023849

Figure 6. Modulation of CDK1 protein level and activity and sensitivity of NSCLC cells to Paclitaxel treatment.

Figure 6

A. CDK1 protein over or under expressed in H157 cells. Cells were transfected with CDK1 plasmid or siRNA, 48 h or 72 h later, were treated with 1 µM paclitaxel for 2 days. CDK1 protein level was monitored by CDK1 or p-Y15 CDK1 antibodies. Sub-G1 DNA content was determined by flow cytometry. B. Knockdown of CDK1 in A549 cells. Cells were transfected with CDK1 siRNA, 72 h later, treated with 1 µM paclitaxel for 2 days. Apoptosis was determined by PARP cleavage using immunoblotting or sub-G1 DNA content using flow cytometry. CDK1 protein level was monitored by CDK1 or p-Y15 CDK1 antibodies. C. Overexpression of CDK1 or CDK1DN in A549 cells. Cells were transfected with CDK1 or CDK1DN plasmids, 48 h later, cell cycling stopped by overnight pretreatment of low serum (0.1% FBS) medium with PI3K inhibitor LY294002 (10 µM), then cells were treated with 1 µM paclitaxel in low serum medium containing LY294002 (10 µM, freshly added) for 2 days. Sub-G1 fraction and cell cycle profile were determined by flow cytometry. Over expression of CDK1 and CDK1DN was detected by CDK1 antibody.