Cell lines were purchased from the American Type Culture Collection (USA) and cultured under 37°C and 5% CO2 in an appropriate medium supplemented with 10% fetal calf serum (FCS), penicillin (100 U/ml), and streptomycin (100 μg/ml). A375 was maintained in Dulbecco’s modified Eagle’s medium, U-2 OS in McCoy’s 5A (modified) medium, A549 in F-12K medium, MCF7 and 769-P in RPMI 1640 medium, and HepG2 in minimum essential medium. To generate fluorescent reporter cells for live-cell imaging analysis of p53 dynamics, we infected the individual cell lines with lentiviruses encoding an established p53-Venus reporter and selected isogenic clones that exhibited dose responses most similar to their respective parental line for conducting the study. The p53-Venus reporter construct, consisting of wild-type p53 fused to a yellow fluorescent protein, Venus, was a gift from G. Lahav at the Department of Systems Biology, Harvard Medical School.

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