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. 2007 Jul 19;26(15):3545–3557. doi: 10.1038/sj.emboj.7601797

Figure 8.

Figure 8

Role of Mediator binding in MED25 function. (A) HeLa cells were transfected with the expression vectors for full-length MED25 (FL) or a MED25 fragment (residues 395–745 or 1–394) with RARE-tk-luciferase in the absence or presence of ligand (1 μM). The fold activation is a comparison between the luciferase activity in the presence and absence of ligand. (B) Flag-MED25 FL or a MED25 fragment (residues 395–745 or 1–394) was expressed in HeLa cells. The interaction was determined by IP using anti-Flag antibody and WB using anti-CBP, anti-TRAP220, and anti-MED6 antibodies. (C) After the transfections described above, crosslinked chromatin was prepared and immunoprecipitated with the antibodies indicated on the right. The precipitates were subjected to PCR analysis using primer pairs spanning the human RARβ2 promoter. (D) HeLa cells were treated with MED25 siRNA (−, untreated; C, control siRNA) and the extracts were subjected to WB with the indicated antibodies. The interaction was determined by IP followed by WB using the indicated antibodies.