Immunoblotting Workflow for Protein Detection

Immunoblotting (IB) was performed as we previously described [25] (link), [26] (link), [27] (link), [28] (link). Briefly, the Blot Mini Gel Tank and Blot Module Set (Thermo) with precast polyacrylamide gels of NuPAGE 10% Bis−Tris protein gels (1.5 mm × 10 wells, Thermo) were used. Primary antibodies included rabbit anti-ISG15 mAb (Sinobiological, Beijing, China; 1:2000), rabbit anti-syntenin mAb (Abcam, Shanghai, China; 1:1000), rabbit anti-TSG101 mAb (Merck, Guangzhou, China; 1:2000), rabbit anti-CD9 pAb (Abcam; 1:2000), and rabbit anti-GRP78/BIP pAb (Proteintech, Wuhan, China; 1:2000). HRP-conjugated anti-rabbit IgG mAb (CST; 1:20000) was used as the secondary antibody.