Gtx629633

The antibodies for NP, GAPDH, CD63, CD81, TSG101 and HSP70 were purchased from Genetex (no. GTX629633, GTX100118, GTX17441, GTX31831, GTX10255 and GTX11573). The antibodies for HA and calnexin were purchased from Millipore (no. 04–902 and AB2301). Cell lysates were prepared using M-PER mammalian protein extraction reagent (Thermo Scientific) with additional protease inhibitors, subjected to electrophoresis on a SDS-PAGE, and transferred onto a Hybond-P membrane. The membrane was probed with the indicated primary and appropriate secondary antibodies, detected using an enhanced chemiluminescence detection kit, and then imaged by Image-Quant LAS4000.

For Western blotting, rabbit anti-ACE2, rabbit anti-IAV nucleoprotein (NP), and goat anti-rabbit IgG antibodies were purchased from GeneTex (GTX101395, GTX629633, and GTX213110-01). Rabbit anti-HA tag antibody was purchased from Millipore (04-902). Mouse anti-IAV matrix protein (M1) antibody was purchased from AbD Serotec (MCA401). For immunofluorescence staining, a mouse anti-IAV NP antibody was purchased from Abcam (ab20343). Alexa Fluor-conjugated secondary antibodies used for immunofluorescence were procured from Molecular Probes (Invitrogen). DAPI was purchased from Sigma-Aldrich.