Tyro3

Samples of treated cells, exosomes and MVs fraction were lysed in RIPA buffer(Santa Cruz, USA) for 20 min on ice and cleared lysate was collected by centrifugation for protein separation on 10% SDS-PAGE gels(Bio-Rad, USA). The proteins were transferred onto Immobilon-FL PVDF Transfer Membrane(Millipore) and detected with appropriate primary antibodies at 4°C overnight, followed by incubation with HRP-conjugated anti-rabbit IgG or anti-mouse IgG (Southern Biotech) secondary antibodies. Membranes were exposed using an enhanced chemoluminescence(NcmECL Ultra) system(NCM Biotech, Suzhou, China) and band intensities were quantified by ImageJ software(NIH). Primary antibodies against CD63(BD Biosciences), CD9(Sigma, #SAB4503606), DAF(#31759), CVB3 VP1(Dako,#M706401-2), Annexin A1(#32934), Annexin A5(#8555), GM130(#12480), Calnexin(#2679) (all from Cell Signaling Technology, USA), CAR(#ab272711), Alix(#ab186429), SCARB2(#ab240186), Tyro3(#ab109231), GAPDH(#ab8245)(all from Abcam, USA), CAV16 VP1(GTX132338), CAV16 3AB(GTX132344), Zika Envelope(GTX133314), Zika Capsid(GTX133317)(All from GeneTex, Inc. Taiwan, China).

Commercially available antibodies were purchased for immunoblotting or immunohistochemical detection of AXL. pAXL Y702, AKT, pAKT S473, ERK, and pERK T202/T204 (Cell Signaling Technology, Danvers, MA); MER (GeneTex, Irvine, CA); TYRO3 (Abcam, Cambridge, MA); Ki67 and cleaved caspase 3 (Biocare Medical, Concord, CA); and β-actin–horseradish peroxidase (Santa Cruz Biotechnology, Dallas, TX). Recombinant human GAS6 was purchased from R&D Systems (Minneapolis, MN).