Ab109210

Metformin and berberine were purchased from MUSTBIO technology (Chengdu, China); Compound C was from CALBIOCHEM (Darmstadt, Germany). Hydroxychloroquine (HCQ), rapamycin, and phthalic acid (PHTH) were from Sigma‐Aldrich (CA, USA). Nicotinamide mononucleotide (NMN) and FK866 were from Santa Cruz Biotechnologies (TX, USA). EX527 was from selleck (CA, USA). Antibodies against LC3 (12741), phospho‐ACC (Ser79) (3661), phospho‐mTOR (Ser2448) (5536), phospho‐p70S6K (Thr389) (9206), and p53 (2524) were purchased from Cell Signaling technology (MA, USA). Those against phospho‐AMPKα (Thr172) (ab133448), Cathepsin B (ab30443), NAMPT (ab109210), AMPKα1 (ab32047), and SQSTM1/p62 (3340‐1) were from Abcam (MA, USA). Antibodies against β‐actin and Lamin B were from Proteintech (Beijing, China), anti‐TFEB (A303‐673A‐M) was from Bethyl (TX, USA), FITC‐goat anti‐rabbit IgG was from Invitrogen (CA). The reagent used for cDNA plasmids transfection was Lipofectamine 2000 (Invitrogen, CA), and that for lentivirus‐shRNA plasmids was X‐treme GENE HP (Roche, CA, USA).

The whole cell lysates were harvested via cell lysis buffer and the protein concentration was detected with BCA Protein Assay Kit (Thermo). Then the proteins were undergoing separating by 8–12% gradient gels and transferred to PVDF (Polyvinylidene Fluoride) membranes. Membranes were blocked by blocking buffer and incubated with primary antibodies at 4 °C overnight. At last the membranes were incubated with secondary antibodies at room temperature for 1 h and scanned by infrared imaging system. The following primary antibodies were used: TSG101 (1:1000, ab133586, Abcam), CD81 (1:1000, ab109210, Abcam), HUR (1:1000, ab200342, Abcam), HER-2 (1:1000, ab134182, Abcam), GAPDH (1:1000, 60,004–1-Ig, Proteintech), p62 (1:1000, cat. no. 18420–1-AP), LC3 (1:1000, cat. no. 14600–1-AP), CD63 (1:1000, BD Bioscience, clone H5C6), CD9 (1:1000, Millipore, clone MM2/57), Fibronection (1:1200, ab285285, Abcam).