Figure 3

NF-κB directly binds to the promoter region of miR-34a gene. (a) EMSA was performed with EC109 cells nuclear extracts using probe corresponding to the third κB site (34a3ΚB) in miR-34a promoter region. Competition and supershift assays against p53, p65 and p50 were also performed. (b) Chromatin derived from EC109 cells were immunoprecipitated with anti-p65, p50, p53 and IgG antibodies. Relative enrichment of each transcription factor-bound DNA was detected by qPCR using primers described in the method. Shown were results normalizing to input DNA. (c) CHIP assay were performed with EC109 cells transfected with p65 or control vectors, Relative enrichment of each transcription factor-bound DNA was detected by qPCR using primer 34aΚB3.