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. Author manuscript; available in PMC: 2014 Mar 7.
Published in final edited form as: Mol Cell. 2013 Jan 11;49(5):843–857. doi: 10.1016/j.molcel.2012.12.006

Figure 1. Brd4 Interacts with Select Chromatin Modifiers and Transcription Factors.

Figure 1

(A) Candidate screen of Brd4-interacting proteins by solution pulldown with purified protein (I, input) incubated with (+) or without (-) f:Brd4. IP with α-Brd4 antibody, followed by IB with anti-test protein antibody.

(B) Brd4 interacts with p53 in 293 nuclear extract.

(C) BID and PDID interact directly and independently with GST:p53, not GST. Proteins purified from bacteria or Sf9 as indicated.

(D) p53 interacts with Brd4 FL, BID and PDID in vivo by BiFC live-cell imaging. Venus-N-p53 and Venus-C-Brd4 containing FL, PDID, BID, PDID-BID, or aa 149-284 were co-expressed in HCT116 p53 -/- cells and imaged by confocal microscopy.

(E) f:PDID (purified from Sf9) and f:BID (purified from bacteria) interact strongly with REG (thick solid line) but weakly with DBD (thin dashed line) of p53. GST pulldown was performed by incubating f:PDID or f:BID with GST or each fusion. Bound f:PDID and f:BID detected by α-FLAG antibody with GST derivatives visualized by CBB staining.

(F) p53 DNA-binding activity inhibited by BID but not PDID. EMSA performed by incubating f:p53, with or without f:PDID or f:BID, with 32P-labeled DNA containing human p21 p53-binding site.