Western blotting was performed as previously described (11 (link)). Cells were washed twice with PBS and lysed with protein lysis buffer (Beyotime, Shanghai, China) containing protease inhibitor cocktail (MCE, USA) and 2% Triton X-100. After 30 min on ice, lysates were treated with 20% loading buffer (Beyotime) for 10 min at 100°C. Proteins were resolved by SDS-PAGE and analyzed by immunoblotting using anti-V5 antibody (1:2,000; Abcam, Cambridge, USA), anti-beta-tubulin antibody (1:1,000; abm, Canada), anti-Flag antibody (1:2,000; Sigma, Burghausen, Germany), anti-gp64 antibody (1:2,000; Abcam, Cambridge, USA), and anti-NPC2 antibody (1:1,000).
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