Fig. 7.

Anti-HBV studies in 3D spheroid models. a Schematic of HBV life cycle indicating the presumed mechanism of action of ETV and CAS9/HBV. b The antiviral drugs used in pre-infected or patient-derived 3D spheroid models. c–e The makers’ expression of HBV in pre-infected 3D spheroid models. c Extracellular HBV-DNA and secreted viral antigens were monitored after drug treatment in pre-infected 3D spheroid models at different time points. Error bars represent s.d. (one-way ANOVA with Dunnett correction for multiple comparisons, n = 4 technical replicates, n.s., non-significant, *P < 0.05, **P < 0.01, ***P < 0.001). d HBsAg staining of pre-infected 3D spheroid models treated with the drugs for 15 days. Scale bars, 25 μm. e QPCR analyses for the expression of cccDNA and 3.5 kb RNA in pre-infected 3D spheroid models treated with the drugs. Error bars represent s.d. (one-way ANOVA with Dunnett correction for multiple comparisons, n = 4 technical replicates, *** P < 0.001). f–h The makers’ expression of HBV in patient-derived 3D spheroid models. f Extracellular HBV-DNA and secreted viral antigens were monitored after drug treatment in patient-derived 3D spheroid models at day 10, day 20 and day 30. Error bars represent s.d.; n = 4 technical replicates. g HBsAg staining of patient-derived 3D spheroid models treated with the drugs for 30 days. Scale bars, 25 μm. h QPCR analyses for the expression of cccDNA and 3.5 kb RNA in patient-derived 3D spheroid models treated with the drugs. Error bars represent s.d.; n = 4 technical replicates