Cholesterol depletion and ceramide enrichment do not reduce cell surface expression of other HCV entry factors. Huh-7w7/mCD81 cells were treated with 7.5 mM MβCD, with 0.5 unit Smase/ml or left untreated (NT) 30 min at 37°C. Cells were then surface biotinylated and lysed in buffer containing 1% Brij97 and divalent ions. Immunoprecipitations were performed with indicated mAbs. Immunoprecipitates were revealed by western blotting using peroxidase-conjugated streptavidin.