Whole cell extracts were prepared in cell lysis buffer [10 mM Tris–HCl (pH 7.4), 1% SDS, and 1 mM Na3VO4]. Western blotting was performed as described previously [27, 30]. Briefly, blots were incubated with primary antibodies diluted 1:500–1:1000 in 5% BSA for 12–16 h at 4 °C, and then incubated with secondary antibody diluted 1:1000–1:2000 in 5% skim milk for 2–3 h at 4 °C. The images were acquired by scanning with the Phosphoimager Typhoon FLA 7000 (GE, Pittsburgh, PA, USA) [27].

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