Figure 2
BST-2 co-localizes with HBs and physically interacts with HBV enveloped particles.
(a) HeLa cells on coverslips, transfected with 500 ng of HBV protein plasmid and 100 ng of BST-2 IHA plasmid, were double-stained with a mouse anti-Flag antibody (mouse anti-myc for HBc) (secondary Ab: Alexa 488) and a rabbit anti-HA antibody (secondary Ab: Alexa 633) and further stained with a PE-conjugated anti-human CD63 antibody. Images were taken under a Zeiss LZM710 confocal microscope. Quantifications of the co-localization were performed using Image-Pro Plus 6.0. At least 20 independent images were examined in each sample and the most representative cells are shown. Scale bars: 20 μm. (b) 293T and Huh-7 cells were co-transfected with 50 ng of BST-2 delGPI IHA and 1 μg of the indicated proviral plasmids. BST-2 and tubulin were examined by Western blotting. The virions were detected by Western blotting for p24CA and by ELISA for HBsAg. (c) 293T cells were co-transfected with 1 μg of HBV proviral, MHBs, or SHBs plasmid, respectively, with or without the BST-2 delGPI IHA plasmid. The clarified culture medium was examined by anti-HA immunoprecipitation. BST-2 was detected by Western blotting and HBsAg by ELISA. (d) The level of immunoprecipitated HBsAg in c) was quantified and shown in columns. These experiments were repeated three times and the most representative data are shown.
