Gm130

Details of western blot could be found in our previous study 21 (link). Briefly, EV samples were lysed using RIPA buffer (Millipore, Bedford, MA, USA) with Protease Inhibitor Cocktail (PIC, final concentration: 35 μg/mL) and PMSF (Phenylmethanesulfonylfluoride, final concentration: 1 mmol/L) for isolation of total proteins. Protein samples (30 μg, concentration determination by BCA kit from Thermo Fisher Scientific) were separated by 8% SDS-PAGE along with a transfer to the polyvinylidene fluoride membrane (Millipore). After blocking with 5% BSA for 1hr at room temperature, the membrane was incubated with primary antibodies overnight at 4 °C and followed by HRP-conjugated secondary antibody for 1 h at room temperature. After interacting with HRP substrate, protein strips were photographed with the ECL detection apparatus (Thermo Fisher Scientific). Primary antibodies for EV characteristic markers CD9 (1:500), CD63 (1:1,000) and TSG101 (1:1,000), and contaminant markers GM130 (1:1,000), albumin (1:500) and calnexin (1:500) (all from System Biosciences, Palo Alto, CA, https://www.systembio.com) were used for analysis by western blotting.