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. 2020 Feb 26;23(3):100867. doi: 10.1016/j.isci.2020.100867

Figure 2.

Figure 2

Sensing and Response to HepG2.2.15.7-Cell-Derived HBV Particles by the α-HBs SNR Cells

(A) Experimental design of α-HBs SNR cells expressing secNL in response to HBV particles and subviral particles derived from HepG2.2.15.7 cells. Purified HBV particles derived from cell supernatants of HepG2.2.15.7 cells were added to α-HBs SNR cells and incubated at 37°C for 48 h followed by measurement of NanoLuc signal.

(B) The NanoLuc activity of α-HBs SNR cells increased in a dose-dependent manner. α-HBs SNR cells were incubated with each amount of HBV particles for 48 h and NanoLuc activity in the cell supernatant measured. Control cells indicate Jurkat T cells harboring only secNL reporter gene without α-HBs SNR. The data shown represent the mean ± standard deviation (SD) of three independent experiments.

(C) Timecourse of NanoLuc activity in the cell supernatant of α-HBs SNR cells incubated with HBV particles. Upper panel indicates the time course of events in this experiment. α-HBs SNR cells were incubated with 0.5 ng/mL HBV particles. Cells were washed and re-suspended in fresh medium every 24 h and NanoLuc activity in the cell supernatant measured prior each medium change. α-HBs SNR cells without exposure to any input stimulus were used as controls. The data shown represent the mean ± SD of three independent experiments. ***p: <0.001. W/MC: wash and medium change.

(D) Timecourse of NanoLuc activity in the cell supernatant of α-HBs SNR cells co-cultured with HepG2.2.15.7 cells. Upper panel indicates the time course of events in this experiment. α-HBs SNR cells were co-cultured with HepG2.2.15.7 cells (HBV-producing cells) or HepG2 cells (uninfected negative control). Cells were washed and medium was refreshed every 24 h and NanoLuc activity in the cell supernatant measured prior each medium change. The data shown represent the mean ± SD of three independent experiments. **p: <0.01, ***p: <0.001. W/MC: wash and medium change.

(E) Response of α-HBs SNR cells incorporated with GFP reporter gene to HBV particles. GFP expressing α-HBs SNR cells were observed by fluorescence microscopy after 48 h of incubation in the presence or absence of 4 ng/mL HBV particles derived from HepG2.2.15.7 cells. Scale bars represent 40 μm.

(F) GFP expression of α-HBs SNR cells increased in a dose-dependent manner. GFP-expressing α-HBs SNR cells were observed by flow cytometry after 48 h incubation in the absence or presence of each amount of HBV particles derived from HepG2.2.15.7 cells. Control cells indicate Jurkat T cells harboring only GFP reporter gene without α-HBs SNR.