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. | Nature Communications

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.

From: The crucial but insufficient role of E2s domain’s residues 490 and 492 in determining the host tropism of hepatitis E virus

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.The alternative text for this image may have been generated using AI.

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.

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