Evidence of WT1 binding in the Bax promoter region in vivo . (A) Schematic showing the Bax promoter and the target region tested in ChIP assay. The locations of primers used to amplify DNA fragments spanning WT1 transcription binding sites were designated in the Bax promoter region. (B) ChIP detection of WT1 transcription factor binding to the Bax promoter region in immature GCs. ChIP assays were performed using DNA extracted from GCs obtained at 36–48 h after transfection. One tenth of the chromatin was kept as input DNA control (Input) before immunoprecipitation. Immunoprecipitations were performed with WT1 antibody, or normal rabbit IgG served as a negative control. DNAs were analyzed by PCR using primers indicated previously. WT1−1299 (210 bp) and WT1−1160 (212 bp) DNA fragments containing WT1 transcription factor binding sites were enriched in chromatin samples immunoprecipitated with WT1 antibody as well as input DNA. A representative gel picture from at least four independent experiments is shown.