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Western blotting Cells were lysed in RIPA lysis buffer [1% NP-40, 0.1% SDS, 50 mM Tris-HCl pH 8, 150 mM NaCl, 0.5% sodium deoxycholate, and Complete EDTA-free protease inhibitor (Roche, 11 873 580 001)] and quantified with the Protein Assay Dye Reagent Concentrate (Bio-Rad, #5000001)