ARV Protein Immunofluorescence Protocol

Methyl-β-cyclodextrin (MβCD), cholesterol, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT), 4′,6-diamidino-2-phenylindole (DAPI) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (Shanghai, China). Filipin III was purchased from Cayman (Michigan, United States). A mouse monoclonal antibody against σC of ARV and polyclonal antibodies against μNS and p10 of ARV were prepared by our laboratory (Duan et al., 2015 (link); Niu et al., 2019 (link)). A rabbit monoclonal antibody against Caveolin-1 was purchased from Cell Signaling Technology (Massachusetts, United States). A mouse monoclonal antibody against β-Actin and secondary antibodies, including fluorescein (FITC)-conjugated goat anti-mouse/rabbit IgG, horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG, and cholera toxin subunit B (CTB) conjugated to Alexa Fluor^® 594 (CTB- Alexa Fluor 594), were purchased from Sigma-Aldrich. RIPA lysis buffer, PMSF, and SDS-PAGE loading buffer were purchased from Beyotime Biotechnology (Shanghai, China). The chemiluminescent substrate and DRAQ5 were purchased from Thermo Fisher Scientific (Shanghai, China).