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. 2003 Jan;23(1):335–348. doi: 10.1128/MCB.23.1.335-348.2003

FIG. 2.

FIG. 2.

Immunoneutralization of Mediator in HNEs inhibits ERα-dependent transcription. (A) Ligand- and p300-dependent transcription by ERα with chromatin templates in vitro. The plasmid template 2ERE-pS2 containing two EREs upstream of the human pS2 promoter (bottom, left) or the plasmid template pERE containing four EREs upstream of the adenovirus E4 promoter (bottom, right) was assembled into chromatin in the presence of recombinant ERα, 17β-estradiol (E2), and p300, as indicated. The chromatin templates were transcribed with an HNE, and the resulting RNA products were analyzed by primer extension. Relative levels of transcription are indicated. (B) Effect of Mediator immunoneutralization on ERα-dependent transcription with chromatin templates. HNEs were incubated with anti-Med220 or anti-Med33 antibodies (40 ng/μl), a combination of both antibodies (20 ng/μl each), or a control antibody (40 ng/μl), as indicated. After the formation of immune complexes, the variously treated HNEs were tested for their ability to support ERα-dependent transcription with chromatin templates as described in panel A.