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. 2007 Apr 2;104(15):6182–6187. doi: 10.1073/pnas.0608717104

Fig. 1.

Fig. 1.

Dependence of transcription on Mediator and TFIIB. (A) In vitro transcription reactions contained Mediator-depleted nuclear extract. Both basal (odd numbered lanes) and HNF-4-driven (even numbered lanes) transcription from a cognate template was monitored. Reactions were supplemented with PC2 (lanes 1 and 2) or increasing amounts of recombinant TFIIB (rTFIIB; 50 ng, lanes 5 and 6; 250 ng, lanes 7 and 8). Control reactions showing residual transcription in Mediator-depleted extract to which neither Mediator nor TFIIB have been added were also included (lanes 3 and 4). (B) In vitro transcription reactions were reconstituted with recombinant TFIIA, TFIIE, TFIIF, and PC4, and affinity-purified TFIID, TFIIH, and Pol II; HNF-4 and PC2 were added as indicated. In addition, reactions contained either no TFIIB (lanes 1–4), 0.4 ng TFIIB (lanes 5–8), or 10 ng TFIIB (lanes 9–12).