Research Paper Volume 12, Issue 20 pp 20702—20727

Transcription factor p53 directly binds to the BAG5 promoter. (A) A schematic diagram of human BAG5 promoter (positions −800 to +200 bps). The putative binding sites of the p53 transcription factor were predicted by ALGGEN-PROMO software. U2OS, HeLa and SH-SY5Y cells were treated with 10 μM of etoposide (B–D) for 48 h or 250 μM of H₂O₂ (E–G) for 24 h. Cell lysates were harvested post 24- or 48-h treatment, and DNA-protein complexes were immunoprecipitated using an anti-p53 antibody or the irrelevant rabbit IgG control. Q-PCR was performed for DNA-protein complexes to detect the DNA fragments at the BAG5 promoter (from -628 to -608, -364 to -344, -219 to -157, and +2 to +18). GAPDH promoter DNA (from -93 to +64) served as a negative control. The amount of immunoprecipitated DNA in each sample is represented as a signal relative to the total amount of input chromatin.