Fig. 7: The critical role of amino acids 490 and 492 in HEV infection in primary porcine hepatocytes in vitro and in swine in vivo.

A Detection of HEV ORF2 protein in supernatants of mutated HEV RNA transfected S10-3 cells. Data represent two biological replicates. B Mutated HEV captured by mAb 6H8 in an immune capture experiment with real time reverse transcription PCR for viral RNA quantitation. Data represent two biological replicates. Cross-genotype mAb 8G12 and an irrelevant mAb (irre.) were used as the positive and negative control, respectively. C Binding capability of the mutant viruses in PPH and HepG2 cells detected by real-time reverse transcription PCR. The dash line indicates the limit of quantitation. Data represent two biological replicates. D The ratio of HEV RNA copies bound to PPH and HepG2 cells. Data represent two biological replicates. E Infectivity of wild-type and mutant viruses in PPH and HepG2 cells detected by ELISA for HEV ORF2 level. Data represent three biological replicates. +A286 and +6H8 represent the infectivity of HEV in the presence of mAb A286 and mAb 6H8, respectively. F Ratio of HEV levels in the supernatant of PPH and HepG2 cells after HEV infection. Data represent three biological replicates. G Number of positive cells detected by immunofluorescence assay after HEV infection. ~69 imaging areas were quantified. Data represent three biological replicates. +A286 and +6H8 represent the number of positive cells detected by immunofluorescence assay after HEV infection in the presence of mAb A286 and mAb 6H8, respectively. H Infection capability of mutant viruses in PPH and HepG2 cells detected by immunofluorescence assay. Green fluorescence indicates HEV ORF2 protein expression and blue fluorescence indicates positive for 4’,6-diamidino-2-phenylindole (DAPI) nuclear staining. Scale bar, 25 μm. I Detection of HEV infection markers in Macaca fascicularis inoculated with wildtype and mutant viruses. A signal to cut-off (S/CO) value of ≥1 was defined as positive. Fecal RNA: HEV RNA in the stool; Serum RNA: HEV RNA in the serum; Ab: anti-HEV antibody. J Detection of HEV infection markers in Bama miniature swine inoculated with wildtype and mutant viruses. A S/CO value of ≥1 was defined as positive. Fecal RNA: HEV RNA in the stool; Serum RNA: HEV RNA in the serum; Ab: anti-HEV antibody. HEV-1: genotype 1 HEV; HEV-4: genotype 4 HEV; PPH: primary porcine hepatocytes. Data are presented as mean values (A–D) or as mean ± SD (E–G). Source data are provided as a Source Data file.