Figure 5.

Effects of PKA and PKC on the association of DAX-1 with the StAR promoter. MA-10 cells were treated without or with increasing doses of (Bu)₂cAMP (0–1.0 mm, A) and PMA (0–20 nm, B) for 6 h, and ChIP assays were carried out as described in Materials and Methods. Cross-linked sheared chromatin obtained from (Bu)₂cAMP and PMA-treated cells was immunoprecipitated (IP) with either IgG or anti-DAX-1 and anti-P-CREB (D) Abs. Recovered chromatin was subjected to PCR analysis using primers (−170/−1 bp, A, B, and D) encompassing the DAX-1 binding site or primers (3596/−3428 bp, C) approximately 3500 bp upstream of the DAX-1 region. Representative autoradiograms illustrate the association of DAX-1 and P-CREB with the StAR promoter in response to (Bu)₂cAMP and PMA. Data shown are representative of three independent experiments.