FIG 5.
DFO induces binding of WT, but not mutant, p53 to Zp. (A and B) Quantitative PCR analysis of chromatin obtained from Sal cells (A) and AGS-Akata cells (B) incubated for 24 h with (+) or without (−) 200 μM DFO prior to immunoprecipitation with a p53-specific or anti-IgG control antibody. (C) Quantitative PCR analysis of chromatin obtained from MutuI cells treated for 24 h with 200 μM DFO prior to immunoprecipitation with the above-mentioned antibodies or a HIF-1α-specific antibody. The cellular p21 promoter and a sequence located 4.8 kbp upstream of Zp served as the positive and negative controls (Neg Con) for p53 binding, respectively. Data shown are means ± SEMs of threshold cycle (CT) values from two independent experiments performed in triplicate. (D) Semiquantitative analysis of chromatin obtained from SNU-719 cells incubated as described for Fig. 1A for 24 h with or without 200 μM DFO prior to immunoprecipitation with antibody specific to p53 or anti-IgG antibody. The analysis was performed by 30 cycles of PCR with primers spanning Zp or a region located 4.8 kbp upstream of Zp as a negative control, followed by electrophoresis in a 1.5% agarose gel.