Fig. 1

Canine liver cells support the full life-cycle of HEV infection. a, In BDE cells upon electroporation of the subgenomic HEV RNA with luciferase reporter, HEV luciferase activity (representing virus replication level) was sustained for 21 days (mean ± SEM; n = 2). b, Upon electroporation of the full-length genomic HEV RNA, BDE cells support long-term production of HEV in the medium for 45 days as quantification of the viral genome by qRT-PCR and calculated as copy numbers (mean ± SEM; n = 2). c. Immunofluorescence staining of viral protein ORF2 (red) in BDE cells. BDE-HEV cells incubated with the matched IgG control antibody serves as negative control, and HEV infected Huh7.5 cells serves as positive control. DAPI (blue) was applied to visualize nuclei. (40× oil immersion objective; Scale bar, 200 μm). d, Anti-HEV activity of ribavirin and interferon-α (IFN-α) in BDE cell based HEV models. Treatment of ribavirin or IFN-α for 24, 48, 72 or 96 h in the subgenomic model determined by luciferase activity (mean ± SEM, n = 5. CTR, non-treatment control.), and in the infectious model determined by viral RNA (mean ± SEM, n = 4. CTR, non-treatment control). *P < .05; **P < .001; ***P < .0001. e, BDE and Huh7.5 cells are permissive for secondary infection with inoculation of BDE cells produced HEV viral particles as determined by qRT-PCR at 24, 48, 72 and 96 h post-inoculation (mean ± SEM; n = 2). The level of HEV infection in Huh7.5 cells for 24 h serves as 1. GAPDH serves as a reference gene