FIG. 7.
Recruitment of ASCOM in RAR transactivation. (A) The retinoid-responsive β-RARE-LUC reporter construct was cotransfected into HeLa cells, along with lacZ expression vector (100 ng) and expression vectors for DN1 (100 ng) or DN1/m (100 ng). Closed and shaded boxes indicate the absence and presence of 0.1 μM 9-cis-retinoic acid (RA), respectively. Normalized luciferase expressions from triplicate samples were calculated relative to the lacZ expressions. Similar results were obtained with CV-1 cells (data not shown). (B and C) ASCOM recruitment to β-RARE and p21WAF1 and H3-K4 methylation. 293T cells were cotransfected with expression vectors for RAR (10 ng), DN1 (100 ng), and DN1/m (100 ng) either in the absence or in the presence of 0.1 μM 9-cis-RA, as indicated. Chromatin from these cells was isolated and immunoprecipitated with the indicated antibodies. The endogenous β-RARE or p21WAF1 region present in the immunoprecipitated samples was amplified by PCR, and input PCR is shown for the loading controls.