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Figure 1


Fig. 1. The PCNA-binding domain of p21 is not essential to block cell-cycle progression. (A) U2OS cells were transfected with GFP-PCNA and the indicated p21 plasmids and the cell-cycle profile of the transfected population was determined. (B) U2OS cells were transfected as in A. BrdU (10 µM) was added 30 minutes before fixation and was detected with specific antibodies. Representative images are shown. The percentage of GFP-PCNA-positive cells that incorporated BrdU was determined (bar chart). At least 200 transfected nuclei/sample were counted. Values are the average and error bars are the standard deviation between equivalent samples in two independent experiments. (C) U2OS cells were transfected as in A, and the sub-nuclear distribution of GFP-PCNA was determined. In the case of the empty vector (EV) and 6Mycp21 (CDK–), ~60% of the cells showed a diffuse distribution, whereas the remainder showed focal PCNA. The two images shown for EV and 6Mycp21 (CDK–) are representative of each situation (merged panels in supplementary material Fig. S5). The quantification of three independent experiments is reported in Fig. 3B.





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