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. 2014 Feb 3;111(7):2500–2505. doi: 10.1073/pnas.1307525111

Fig. 1.

Fig. 1.

Genetic regulation of Med3 target gene expression. Total RNA was isolated, and FLO11 expression levels were determined using qRT-PCR. (A) MED3 and SPT8 are required for CDK8-mediated FLO11 derepression. (B) CDK8 suppresses the activation function of MED3. Strains carrying plasmids expressing MED3 or MED2 (3) together with CDK8 or CDK8 (D290A) (29) were maintained in appropriate selective media. Values are given as a percentage relative to either (A) cdk8Δ or (B) MED3 set at 100% and normalized to ACT1. (C) Med3 recruitment to the FLO11 promoter is regulated by CDK8. Chromatin immunoprecipitates were prepared from cells expressing Med3, Cdk8, or Med3 and Cdk8 using anit-HA and then analyzed for the presence of Med3 on the heat shock–like element (−488 to −342 from ATG, white bars) (13), TATA box (−183 to −53 to ATG, black bars), and 3′ ORF (3,628–3,765 to ATG, hashed bars) of FLO11 using qRTPCR. 0, no antibody control.