Western Blot Analysis of Protein Expression

Western blot analysis was performed as described previously [34 (link)]. Briefly, cells were washed three times with PBS, lysed in lysis buffer (50 mM Tris–HCl [pH 6.8], 1% sodium dodecyl sulfate, 10% glycerol), and stored at − 70 °C until use. Equal amounts of total proteins were separated by SDS–polyacrylamide gel electrophoresis, transferred to a polyvinylidene fluoride membrane (Millipore), and proteins were immunodetected using the appropriate primary antibody followed by incubation with horseradish peroxidase-conjugated secondary antibody. Amersham ECL Prime western blotting Detection Reagent (GE Healthcare Life Sciences) was used to visualize antibody-antigen complexes. All used antibodies are listed in Table S2.