FIGURE 3.

PRMT5 and WDR77 interact with purified Mediator complexes. A and B, Western blotting of two-step purified HF-CDK8-containing (A) or HF-CDK19-containing (B) Mediator complex. Nuclear extracts from cells stably expressing HF-CDK8 were separated on an HPLC-TSK G4000SW XL gel filtration column. The 2-MDa fractions were further purified using anti-FLAG M2 agarose column or normal mouse IgG-conjugated agarose column. Lane 1, 5% input; lane 2, normal mouse IgG eluate (nIgG); lane 3, M2-agarose eluate (FLAG M2). Each protein indicated on the right side was detected with the corresponding antibody. IP, immunoprecipitation.