Figure 1

p53 inactivation up-regulates IL-6 expression and induces the loss of methylation at IL-6 proximal promoter. a) IL-6 ELISA assay and RT-PCR analysis of IL-6 mRNA level in MCF-7 and MDA-MB231 cells; b) quantitative evaluation of IL-6prox methylation-specific PCR analysis (MS-PCR) in MCF-7 and MDA-MB231 cells. "U" stands for Unmethylated and "M" Methylated specific PCR product; c) Western blot analysis of p53 protein level, RT-PCR analysis of Bnip3 and p21Waf1 mRNA level and luciferase assay of p53 responsive reporter plasmid (p53-Luc) in MCF-7 cells transduced with empty (pBabe) or p53 mini-protein (p53D) retroviral vector; d) IL-6 ELISA assay, RT-PCR analysis of IL-6 mRNA level and quantitative evaluation of IL-6prox MS-PCR analysis in pBabe/p53 D cells. β-Actin was assessed as quantitative control for RT-PCR and Western Blot. Student t test, *p < 0.05, ^#p < 0.005. NSF: Non Specific Fragment