FIG. 5.
Pools of GM00011 cells were generated carrying vector alone, E6, the dominant-negative p53 mutant, or E7. Cells were either left untreated (No UV) or exposed to UV light at a dose of 10 J/m2 and harvested 4 h (+UV 4 h) or 8 h (+UV 8 h) later. (A) Levels of p53 were determined by Western analysis as described in the legend to Fig. 1. The first lane contains 10 μl of a rabbit reticulocyte lysate programmed with p53 RNA (IVT, in vitro translated). The second lane contains 2 μl of a rabbit reticulocyte lysate programmed with p53 RNA. The next 12 lanes contain 150-μg quantities of lysates from NDFs infected with the retrovirus indicated. (B) Levels of mdm2 mRNAs were analyzed by S1 nuclease protection. The ratio of the amount of mdm2 mRNA synthesized from the P2 (p53-responsive) promoter to the amount of mdm2 mRNA synthesized from the P1 (constitutive) promoter is shown. The results are averages of data from two experiments with two independent sets of pools of infected NDFs. One of the sets of NDFs was also used to generate the data shown in Fig. 5A.