Immunoblotting Technique for Protein Analysis

Immunoblotting assays were performed as previously described [18 (link)]. SDS-PAGE was performed on the extracted proteins, which were then transferred onto PVDF membranes (BIO-RAD, Hercules, CA, USA). The gel bands formed after exposure to appropriate antibodies (primary and secondary) were visualized using the ECL reagent (BIO-RAD) and imaged by Tanon 5200-Multi (Tanon Science & Technology, Shanghai, China). The primary antibodies (against TFEB, GFP, LAMP2, NDRG1, SMPD1, and actin) and the HRP-conjugated secondary antibodies (goat anti-rabbit antibody) were supplied by Proteintech (Wuhan, China). Original uncropped images for immunoblotting are presented in Figure S1.