Figure 3

Recognition of TEM-associated CD81 in Huh-7w7/mCD81 cells. A, Flow cytometry analysis of Huh-7w7/mCD81 cells stained with the mAbs MT81 and MT81w. Cells stained only with PE-conjugated secondary antibody were used as control (dotted line). B, Cell lines were surface biotinylated and lysed in the presence of Brij97 or Triton X-100 before immunoprecipitation with MT81, MT81w and 5A6 mAbs. Immunoprecipitates were revealed by western blotting using peroxidase-conjugated streptavidin. C, Cell lines were surface biotinylated and lysed in buffer containing 1% Brij97 and divalent ions. Immunoprecipitations were then performed with 5A6, MT81, MT81w, 8A12 (anti-EWI-2), TS151 (anti-CD151) or irrelevant (CTL) mAbs. Immunoprecipitates were revealed by western blotting using peroxidase-conjugated streptavidin. The molecular weights of the prestained molecular ladders are indicated in KDa. The asterisks indicate dimers of CD81.