Coimmunoprecipitation was carried out as described (21). Briefly, 25 μg of extracts from wild-type or Brpf1−/− MEFs (E13.5) was preincubated with 5 μg of antibody in the NP-40 buffer [50 mM tris-HCl (pH 8.0), 50 mM NaCl, 1% NP-40, 50 mM NaF, and protease inhibitor cocktail] with overnight rotation at 4°C. Immunoprecipitation was then performed by incubating each mixture with 25 μl of Dynabeads Protein G (Invitrogen, 10003D) with rotation for 45 min at 4°C. After washing four times with 0.5 ml of PBS containing 0.02% Tween 20 and protease inhibitor cocktail, the Dynabeads were resuspended in the reducing SDS sample buffer and boiled for 5 min. The supernatant was used for SDS-PAGE and immunoblotting analysis with the indicated antibodies.

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