FIG. 7.
TBP-associated proteins. (A) Western blot analysis of eluate from TBP affinity column eluates. Crude cell extract was passed over GST or GST-TBP columns. Bound proteins were eluted with 2 M KCl and probed with antibodies against Not1 and TafII90. Lanes indicated are onput (O), flowthrough (F), wash (W), and eluate (E). (B) Western blotting of immunoprecipitates from crude fractions of whole-cell extract derived from cells with (FLAG-TBP) or without (untagged) FLAG epitope-tagged TBP reveals that Not proteins interact, directly or indirectly, with TBP. Under these conditions, TBP also interacts with other proteins previously described as TBP-interacting factors, including TafII90, NC2, Spt3, and Mot1, but not RNA polymerase II. Lanes indicated are onput (O), flowthrough (F), wash (W), and eluate (E).