Cells were harvested and the proteins were extracted. Western blot analysis was performed as previously described [25]. Briefly, proteins were resolved on SDS-PAGE, transferred onto polyvinylidenedifluoride transfer membranes (PVDF), and blocked with 5% non-fat dry milk in TBST buffer for 1 h at room temperature. The membranes were then incubated with the diluted primary antibodies overnight at 4℃, washed three times, followed by the incubation with HRP-conjugated secondary antibodies for 1 h at room temperature. The bands were detected with an EZ-ECL kit (BI Biological Industries, 20-500-120) in the MicroChemi bio-imaging system (DNR).
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