FIGURE 2. Association of transcription factors to MnSOD promoter and enhancer element.
The binding of transcription factors to the promoter or enhancer region of MnSOD gene was evaluated by ChIP assay in HepG2 cells as described under “Experimental Procedures.” A, chromatins were immunoprecipitated by Sp1 and p50 antibodies or IgG. The pull-down proteins in the immunoprecipitates were detected by Western analysis. DMSO, Me2SO. B, immunoprecipitated DNA was amplified by PCR using primers targeted to the enhancer element (1742–2083) and the promoter elements (−154 to −6) of the MnSOD gene. kb, kilobases. C, the immunoprecipitated DNA was amplified by PCR using primer specific to an off-target region on MnSOD gene (+521 to +839). Total genomic DNA obtained from HepG2 cells were parallel-amplified by using the same off-target primer as input DNA control.
