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. 2015 Nov 23;5:17047. doi: 10.1038/srep17047

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Copyright © 2015, Macmillan Publishers Limited

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(a) A cell suspension of the HepG2/NTCPA3 cell line was prepared with EDTA only or with both trypsin and EDTA. These suspended cells were infected with HBV at 1,000 GEq/cell. The resulting cells were harvested at the indicated time. The amount of intracellular HBV RNA was estimated by real-time qRT-PCR. Data were calculated from 4 independent experiments and are expressed as means ± SD. (b) HepG2 cells (lanes 1 and 2) or HepG2/NTCPA3 cells (lanes 3 and 4) were collected using a cell scraper (lanes 1 and 3) or by trypsin-EDTA treatment (lanes 2 and 4). The membrane fractions were prepared from the harvested cells and then incubated with (+) or without (−) PNGase for deglycosylation. These membrane proteins were subjected to Western blot analysis using an antibody to NTCP (upper panels) and CBB staining (lower panels).