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. Author manuscript; available in PMC: 2021 Sep 26.
Published in final edited form as: Cell Microbiol. 2020 Sep 28;22(12):e13250. doi: 10.1111/cmi.13250

Figure 2. Quantifying HBV attachment to target cells.

Figure 2

(a) HBV attachment to HepG2 cells is NTCP independent. HepG2-NTCP K7 and Huh7-NTCP along with parental cells were stained for NTCP expression using Atto488 labelled Myrcludex B (200 nM) and imaged using 63× objective (scale bars indicate 20 μm). (b) Pre-chilled HepG2, HepG2-NTCP K7 cells, Huh-7 and Huh7-NTCP were inoculated with HBV (MOI 200) for 1 h on ice, unbound virus removed by washing and cells treated with trypsin or left untreated and cell-associated HBV DNA quantified by RT-PCR. Data is expressed relative untreated HepG2-NTCP cells. (c) HBV attachment to HepG2 cells is dose-dependent. Increasing dose of HBV (MOI 20-2000) was inoculated with HepG2-NTCP K7 and HepG2 cells for 1 h at 4°C, unbound virus removed by washing and cell-associated HBV DNA quantified. HBV DNA levels are expressed relative to PRNP and represent two independent experiments presented as mean ± standard error of the mean (SEM). Each experiment consisted of three replicates per condition. Statistical analysis was performed using a Mann–Whitney U test (*p < .05, **p < .01, ***p < .001)