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

Figure 1.

Figure 1

Establishment of HBV-Sensing Cells Using Anti-HBs scFv synNotch Receptor (α-HBs SNR) and Sensing HBs Protein by α-HBs SNR Cells

For a Figure360 author presentation of this figure, see https://doi.org/10.1016/j.isci.2020.100867.

(A) Schematic diagram of α-HBs SNR cells against HBsAg. The α-HBs SNR cell has α-HBs SNR on cell surface and Gal4-response gene in the nucleus. SynNotch receptor has myc-tagged scFv for HBsAg binding, the Notch core activation domain, and an artificial transcription factor (TF), Gal4-VP64. Reporter genes (secreted NanoLuc luciferase (secNL) or GFP) are designed to be regulated by Gal4 TF via an upstream activating sequence (UAS).

(B) The α-HBs SNR cells expressed secreted NanoLuc luciferase (secNL) in response to liposomal recombinant LHBsAg (rLHBsAg) in dose-dependent manner. The control or α-HBs SNR cells and rLHBsAg were incubated at 37°C, and the NanoLuc activity in the cell supernatant was measured after 48 h. Control cells indicate Jurkat T cells harboring only secNL reporter gene without α-HBs SNR. Each value is normalized to those obtained from without rLHBsAg and represents the mean ± standard deviation (SD) of three independent experiments. Horizontal lines represent axis break of the y-axis. **p: <0.01, ***p: <0.001.

(C) The α-HBs SNR cells sensed HBsAg secreted by Tet-ON-SHBs cells in dose-dependent manner. α-HBs SNR cells and HBsAg derived from untreated or 5μg/mL doxycycline (Dox)-treated Tet-ON-SHBs cells were incubated at 48 h and then the NanoLuc signal in the supernatant was measured. Control cells indicate Jurkat T cells harboring only secNL reporter gene without α-HBs SNR. Amount of HBsAg was calculated using HBs ELISA kit. Each value is normalized to those obtained from without HBsAg and represents the mean ± SD of three independent experiments. *p: <0.05, ***p: <0.001.