Cellular lysates in radioimmunoprecipitation assay (RIPA) buffer containing a protease inhibitor cocktail (Sigma-Aldrich) were normalized by the total protein level and analyzed by SDS-PAGE and immunoblotting with corresponding antibodies. For immunoprecipitation, precleared cytosolic fractions were incubated overnight with 2 μg of corresponding antibodies at 4°C, followed by 1-hour incubation with Protein A/G Agarose (Santa Cruz Biotechnology) (50 μl of a 50% slurry). Resin was washed three to five times, and pulled-down proteins were analyzed by SDS-PAGE and Western blotting with corresponding antibodies.

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