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. 2006 Jan 24;34(2):543–554. doi: 10.1093/nar/gkj460

Figure 2.

Figure 2

Location of p53 protein within chromatin on human p21(waf1/cip1) promoter. Nineteen overlapping regions evenly cover the first 7.1 kb of the human p21(waf1/cip1) promoter (A) Due to IgG binding to region 12 an alternative region 12C was chosen (Table 1). Chromatin was extracted from MCF-7 cells that had been treated for 360 min with solvent (DMSO) or 300 µM fluorouracil. ChIP assays using an antibody against p53 were performed (B) Precipitations with IgG and anti-GST antibody served as controls for the specificity of the p53 detection. Representative agarose gels of the input lane confirm the comparable detection sensitivity of the 20 different promoter regions. The induction of p53 binding by 5-fluorouracil to promoter regions 1, 5, 7 and 12C was determined by quantitative real-time PCR (C) Columns indicate the means of at least three independent cell treatments and the bars represent standard deviations. A one-tailed Student's t-tests was performed to determine the significance of the increase in p53 binding in fluorouracil-treated samples in reference to solvent controls (*P < 0.05, **P < 0.01).