Western Blotting Protocol for RPMS1 Detection

Western blotting was performed as described previously [30 (link)]. Briefly, cells were lysed in mammalian cell lysis buffer, and proteins within the clarified lysates were resolved by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes for immunoblotting against the corresponding antibody. The results were revealed using enhanced chemiluminescent (ECL) detection reagents (Beyotime Co., Shanghai, China). The rabbit polyclonal anti-RPMS1 antibody was from Proteintech Group Inc. (Wuhan, Hubei, China), and the human anti-β-actin antibody was from Sigma-Aldrich Co. (St. Louis, MO, USA). A horseradish peroxidase (HRP)-conjugated anti-rabbit IgG antibody was used as the secondary antibody (Promega, Madison, WI, USA).