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. 2004 Oct;24(20):9059–9069. doi: 10.1128/MCB.24.20.9059-9069.2004

FIG. 2.

FIG. 2.

Brd4-SPA-1 interaction in vivo. (A) Coimmunoprecipitation of endogenous Brd4 and SPA-1. Extracts from HeLa cells were immunoprecipitated with the antibodies indicated on the top and tested for Brd4 and SPA-1 by immunoblotting. Input represents 2% of extracts. (B) Coimmunoprecipitation of ectopically expressed Brd4 or SPA-1. Extracts from HeLa cells transfected with Flag-Brd4, GFP-SPA-1, or control empty vectors were precipitated with anti-Flag or GFP antibody and blotted with the antibody indicated on the left (the band corresponding to GFP alone produced from control vector was not included in the figure). Input represents 2 or 4% of the extracts used for precipitation. (C) Molecular mass estimation of the Brd4 complex. Q-Sepharose-purified nuclear extracts (500 μg) of the 0.3 M KCl-Q-Sepharose fraction were subjected to gel filtration analysis on a Superdex 200 column by use of the SMART system (Pharmacia Biotech). Fractions were resolved on an SDS-10% polyacrylamide gel and analyzed by immunoblotting. The estimated molecular mass is shown on top.