Ab285285

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).

The serum levels of testosterone (ab285350, Abcam, UK), estradiol (E2, ab285285, Abcam), luteinizing hormone (LH, CEA441Ra, USCN Life Science and Technology Company, Wuhan, China), follicle-stimulating hormone (FSH, CEA830Ra, USCN), and fasting insulin (FINS, ab273188, Abcam) were detected using commercial ELISA kits following the manufacturer’s protocols. Fasting plasma glucose (FPG) level was assessed with a blood glucose meter (Omron, USA). Homeostasis model assessment of insulin resistance (HOMA-IR) = FINS × FPG/22.5.