FIG. 2.

Association of dMED6 with Mediator complex. (A) Coimmunoprecipitation of dMED6 with other Mediator subunits. Drosophila embryo nuclear extract was immunoprecipitated with anti-dSOH1 Ab, and the input (I), supernatant (S), and pellet (P) were analyzed by immunoblotting with Abs against the proteins indicated at the left. (B) Gel filtration analysis of dMED6-containing complex. Embryo nuclear extract was put on a Superose-6 gel filtration column, and the filtrates were analyzed with Abs against the proteins indicated at the left. The input and the elution positions of size markers are marked. (C) Transcriptional activation of the E4 promoter constructs by Gal4-VP16 in nuclear extracts. Before the in vitro transcription assay, the nuclear extracts were immunodepleted with anti-dSOH1 (α-dSOH1) or anti-β-galactosidase (mock). Recombinant Gal4-VP16 (40 ng) was added to the reaction mixtures as indicated. Arrows, transcripts from the E4 templates containing five tandem Gal4 DNA binding sites (G5-E4).