FIG. 1.

(A) Resolution of two different RNAPII complexes by Sepharose CL-4B gel filtration chromatography. The upper panel summarizes the Western blot results shown below. The lower panel shows Western blots of fractions of the Sepharose CL-4B gel filtration column. Blots were probed with antibodies as indicated at the side of each blot. IIF (RAP74), the RAP74 subunit of TFIIF cycC, cyclin C. Input (1%; IN) and fraction numbers are indicated on the top of the lower panel. The last row shows the transcription activity of the column fractions obtained by using the adenovirus major late promoter in a system reconstituted with TBP, TFIIB, and TFIIH. (B) The amount of acetate incorporated into core histones and the protein concentration of each Mono S (see Materials and Methods) column fraction. (C) Coelution of CBP, PCAF, RNAPII (RPB1), BAF47, cyclin C, and CDK8 by Mono S ion exchange chromatography. Fractions from a similar Mono S column were analyzed by Western blotting using antibodies against CBP, PCAF, RPB1, BAF47, cyclin C, and CDK8.