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Protein isolation and western blotting
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Cell pellets were lysed in RIPA buffer (150 mM NaCl; 50 mM Tris pH 8.0; 1.0% Igepal CA-630; 0.5% sodium deoxycholate; 0.1% SDS; Sigma-Aldrich) supplemented with protease inhibitors (cOmplete mini, Roche Diagnostics; 1 mM PMSF; 10 mM NaF; 2.5 mM Na3VO4) and centrifuged at 10,000 × g at 4 °C for 30 min. The resulting supernatant was then assessed by standard immunoblotting.
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