FIG. 5.

p53-dependent regulation of representative candidate target gene expression. The isogenic pair of HCT116 p53^+/+ and p53^−/− cells were treated with ADR (350 nM) for 0, 6, 12, and 24 h; the HIp53 cells (p53) and corresponding vector control cell line (Ø) were treated with ponasterone A (10 μM) for 24 h, and the HK cells were infected with a GFP- or p53-expressing adenovirus for 30 h. Total RNA from HCT116 cells and mRNA from HIp53 and HK cells was purified and reverse transcribed and quantitative real-time PCR performed. The samples were normalized to GAPDH, and the results are presented as changes relative to either the 0-h HCT116 p53^+/+ sample (left panel), the HIp53 vector control cells treated with ponasterone (middle panel), or the HK cells infected with a GFP-expressing adenovirus (right panel). The results are the means of three independent experiments (MCF-10A cells and HMEC) or duplicate experiments (HK cells), with error bars representing the standard deviations. Note that the y axes are set to 7.0 with the exceptions of the left and middle panels for the CDKN1A gene and the left and right panels for the EDN-2 gene.