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. Author manuscript; available in PMC: 2018 Oct 27.
Published in final edited form as: Science. 2018 Jun 21;361(6400):eaar3958. doi: 10.1126/science.aar3958

Fig. 4. Intrinsically disordered regions (IDRs) of BRD4 and MED1 phase separate in vitro.

Fig. 4.

(A) Graphs plotting intrinsic disorder (PONDR VSL2) for BRD4 and MED1. PONDR VSL2 score (y-axis) and amino acid position (x-axis) are shown. Purple bar designates the IDR under investigation. (B) Schematic of recombinant mEGFP fusion proteins used here. Purple boxes indicate IDR’s of BRD4 (BRD4-IDR) and MED1 (MED1-IDR) shown in (A). (C) Visualization of turbidity associated with droplet formation. Tubes containing BRD4-IDR (left pair), MED1-IDR (middle pair) or GFP (right pair) in the presence (+) or absence (−) of PEG-8000 are shown. Blank tubes included between pairs for contrast. (D) Representative images of droplet formation at different protein concentrations. BRD4-IDR, MED1-IDR or mEGFP were added to droplet formation buffer to final concentrations indicated. (E) Representative images of droplet formation at different salt concentrations. BRD4-IDR or MED1-IDR was added to droplet formation buffer to achieve 10 μM protein concentration with a final NaCl concentration as indicated. (F) Representative images of droplet reversibility experiment. BRD4-IDR (top row) or MED1-IDR (bottom row) BRD4-IDR or MED1-IDR, as indicated, (20 μM protein, 75 mM NaCl) (initial) or followed by a 1:1 dilution (diluted 1/2) or a 1:1 dilution with an increase to 425mM NaCl (diluted 1/2 + NaCl)