Skip to main content
. 2012 Jan;32(1):26–35. doi: 10.1128/MCB.05513-11

Fig 2.

Fig 2

PNR-mediated stimulation of p53-responsive promoters requires p53. (A) In p53-null H1299 cells, PNR alone fails to stimulate p53RE-FLuc whereas PNR in the presence of exogenous p53 stimulates p53RE-FLuc. The RLU in the sample with 0 μg of p53 and PNR was normalized to 1. (B) shRNAs targeting p53 abolish PNR-mediated stimulation of p53RE-FLuc in HeLa cells. shRNA-GFP-expressed shRNA targeting GFP was used as a control. The RLU in the sample with both shRNA-GFP and PNR was normalized to 100%. Upper panel: immunoblot showing β-actin loading control and shRNA-mediated reductions in p53 protein accumulation. The numbers below each band represent the level of p53 protein normalized to β-actin and to the p53 level in sample 1. Lower panel: reporter assay. (C) shRNAs targeting p53 significantly inhibit PNR-mediated increases of p21 mRNA levels in HeLa cells. The p21 mRNA level was measured by qRT-PCR normalized to the sample with only shRNA-GFP transfected. (D) Mutation of p53-responsive elements abrogates PNR-mediated stimulation of MT-p53-RE-FLuc in HeLa cells. The RLU in the sample with only p53RE-FLuc was normalized to 1. (E) Dominant-negative mutant p53-C135Y abrogates the PNR-mediated stimulation of p53RE-FLuc in HeLa cells. The RLU in the sample with only p53RE-FLuc was normalized to 1.