Buffer A contained 50 mM tris-HCl (pH 7.5) and 0.1 mM EGTA. Lysis buffer was 50 mM tris-HCl (pH 7.5), 1 mM EGTA, 1 mM EDTA, 50 mM sodium fluoride, 5 mM sodium pyrophosphate, 1 mM sodium orthovanadate, 1% (w/v) Triton X-100, 0.27 M sucrose, 0.1% (v/v) 2-mercaptoethanol, and protease inhibitors (complete protease inhibitor cocktail tablets, Roche, 1 tablet per 50 ml). TBS-Tween buffer (TTBS) was tris-HCl (pH 7.5), 0.15 M NaCl, and 0.2% (v/v) Tween 20. SDS sample buffer was 1× NuPAGE LDS sample buffer (Invitrogen) containing 1% (v/v) 2-mercaptoethanol. Protein concentrations were determined following centrifugation of the lysate at 16,000g at 4°C for 20 min using the Bradford method with bovine serum albumin as the standard.

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