Western Blot Protein Detection

Western blotting assay was performed according to the previous studies (Zhong et al., 2012b (link); Quan et al., 2015 (link)). Briefly, cells were harvested and the total proteins were extracted with RIPA lysis buffer after the required treatments. Equal amounts of total proteins were separated by appropriate SDS-PAGE followed by transferring onto a PVDF membrane. After blocking with non-fat milk, the membrane was incubated with specific primary antibodies and the corresponding second antibodies, respectively. The specific protein bands were visualized with an Amersham^TM ECL^TM advanced western blotting detection kit (GE Healthcare Life Sciences, United Kingdom).