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Figure 7


Fig. 7. p21-PCNA interaction is detrimental for cell survival after UV irradiation. (A) U2OS cells transfected with empty vector (EV) or 6Mycp21 (CDK–) were UV irradiated (10J/m2). After fixation, immunostaining with phosphorylated H2AX ({gamma}H2AX)- and p21-specific antibodies was performed. DAPI staining was used to visualize the nucleus. A complete panel showing all p21 mutants is shown in supplementary material Fig. S5. (B) U2OS cells transfected with the indicated plasmids were irradiated with 10J/m2. The number of cells with {gamma}H2AX accumulation was quantified for each time point. The data shown represent the percentage of p21-positive cells with detectable accumulation of {gamma}H2AX. In all cases, 100 transfected nuclei were counted. The last column in each group corresponds to the percentage of total cells with detectable accumulation of {gamma}H2AX. Values are the average and error bars are the standard deviation between equivalent samples in two independent experiments. (C) U2OS cells transfected with EV or 6Mycp21 (CDK–) and GFP-pol {eta} were UV irradiated utilizing polycarbonate filters and {gamma}H2AX staining used to detect the irradiated areas. (D) U2OS cells transfected with GFP-PCNA and the indicated p21 plasmids were UV irradiated (10J/m2). The cell-cycle profile of the transfected population was determined 48 hours later.





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