Western blot analysis was carried out to assess protein expression levels. Total protein was extracted from the cells by addition of lysis buffer (10 mM Tris, pH 8.0, 120 mM NaCl, 0.5% NP-40, 1 mM EDTA) containing protease inhibitor cocktail (Roche, Switzerland) and phosphatase inhibitor cocktail (Roche). Cultured cells were scraped and whole-cell lysates were prepared by centrifugation at 16,363 × g for 15 min at 4 °C. Equal amounts of protein were loaded alongside a pre-stained protein ladder (1st Base) and electrophoresed on 10% SDS-polyacrylamide gels. The resolved proteins were transferred onto PVDF membranes (Immobilon transfer membrane, Millipore Corporation). After electroblotting, the membranes were blocked with Tris-buffered saline with Tween 20 (1xTBST: 20 mM Tris base, 137 mM NaCl, 0.1% Tween 20) containing 5% non-fat dry milk at room temperature for 1 h before overnight hybridization with appropriate concentrations of primary antibodies (Supplementary Table 3) diluted in blocking buffer. The membranes were then incubated with anti-rabbit HRP-conjugated secondary antibody (NA934, GE Healthcare, UK) for 1 h at room temperature. Antigen-antibody complexed to the blotted proteins were detected using chemiluminescence detection system. The chemiluminescent signal was developed on Amersham Hyperfilm ECL (GE Healthcare).

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