FIG. 2.

GST-rv-cyclin AD pulls down TAF9. A. Pulldowns from 75 μg HeLa nuclear extract by GST protein, by wild-type GST-rv-cyclin AD (GST-240-270) (wt) and mutated forms V260F and V260S, and by GST-VP16C and corresponding mutated form F475A. The Western blot was probed with antibody reactive to the amino terminus of human TAF9 (aTAF9) and reprobed with anti-GST (αGST; bottom panel), which recognizes the input GST and GST fusion proteins. The lane labeled “input” represents 5% of the total extract used for each pulldown. B. Alignment of TAF9-binding motifs identified by Choi et al. (2) with the VP16C AD (aa 455 to 486) and rv-cyclin AD (aa 240 to 270). The relative amino acid position of each motif within its respective protein is indicated after each name. Similar and identical residues are shaded. An asterisk indicates the relative positions of VP16C F475 and rv-cyclin V260. ALL-1, acute lymphocytic leukemia protein; HSF-1, heat shock transcription factor 1; ESX, epithelial restricted with serine box, also known as E74-like factor 3 [ELF3] and epithelial-specific Ets protein; NFAT1, nuclear factor of activated T cells; NFκB p65, nuclear factor kappa-B; NF-IL-6, nuclear factor for interleukin-6 expression; and p53, tumor protein 53.