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. 2011 Aug 11;286(39):33709–33718. doi: 10.1074/jbc.M110.193870

FIGURE 1.

FIGURE 1.

CF1 subunits and poly(A) polymerase copurify with TFIIB on an anti-HA affinity column. A, HA-tagged TFIIB was affinity-purified from cells harboring Myc-tagged CF1 subunits or poly(A) polymerase as described under “Experimental Procedures.” Purified samples were subjected to SDS-PAGE followed by Western blot analysis using anti-HA and anti-Myc antibodies. Lane 1 displays molecular weight (MW) marker proteins, and lane 2 represents Imperial Coomassie Blue staining of the eluate from an anti-HA affinity column. B, affinity purifications were performed for HA-tagged TFIIB in the wild type and the looping-defective mutant sua7-1 strain. The eluates from the affinity columns were subjected to SDS-PAGE, and Western blotting was performed with antibodies against TFIIB, Rna15, and poly(A) polymerase. C, affinity-purified HA-tagged TFIIB was subjected to SDS-PAGE followed by Western blot analysis using anti-HA, anti-TBP, anti-Kin28, and anti-Rpb1 antibodies. IP, immunoprecipitation.