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. 2017 Nov 30;7:16616. doi: 10.1038/s41598-017-16882-5

Figure 3.

Figure 3

HepG2-NTCP cells produce infectious HBV. (A) Schematic of experimental design. (B) Immunofluorescence microscopy to detect HBcAg in infected HepG2-NTCP cells. HepG2-NTCP cells were infected with 100 GEQ/cell of HBV in the presence of 2% DMSO and 4% PEG. One day post-infection fresh medium containing 2% DMSO was added. At 7 dpi supernatant from donor cells was transferred to naïve HepG2-NTCP cells (acceptor). On the day of transfer, naïve cells were treated with 4% PEG, 2% DMSO in the presence of absence of 500 nM MyrB for 24 h. 2% DMSO was included through the entire course of infection. As controls (acceptor) HepG2 cells that do not express NTCP and HepG2-NTCP cells treated with MyrB were used. (C) HepG2-NTCP cells were infected with HBV in the presence of MyrB (donor) and supernatant was transferred on naïve HepG2-NTCP cells (acceptor). On the day of transfer cells were treated with PEG for 24 h. This experiment was used as a control for carryover inoculum. (D) HepG2-NTCP cells were infected with HBV following the same protocol with panel B (donor). One day post-infection ETV was added and kept until 7 dpi. The supernatant was then transferred on naïve HepG2-NTCP cells (acceptor). (E) Quantification of HBeAg levels released in supernatant from acceptor cells. Six biological replicates were performed and data are shown as means ± s.d. (one-way ANOVA *P < 0.001).