Western blot analysis was performed as previously described with minor modification36 (link). Cells were lysed with RIPA buffer (Sigma Aldrich, St. Louis, MO, USA) containing a protease inhibitor cocktail and PhoSTOP (Roche Molecular Biochemicals, Basel, Switzerland). Ten micrograms of the protein lysate were loaded into 10% SDS-PAGE and transferred to PVDF membranes (Amersham, Arlington Heights, IL, USA). After transfer is finished, the membranes were incubated with blocking solution and then appropriate antibodies. The bands were visualized with Advanced ECL Western Blotting Detection Reagents (Amersham). GAPDH levels are used as a loading control.
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