Figure 6

Spicatoside A inhibits replication of the HEV genotype 3 strain 47832c. A549 cells were inoculated with cell culture supernatants containing HEV genotype 3 strain 47832c and incubated at room temperature for 1 h, followed by treatment with either DMSO or spicatoside A. Cells were re-treated with either DMSO or spicatoside A every 3 d after the initial treatment. At 1, 3, 7 and 14 d after inoculation, (A) the relative amounts of HEV RNA in cells and (B) the amounts of HEV RNA copies in culture media were determined via qRT-PCR using primers specific for HEV ORF1. Relative amounts of cell-associated HEV RNA in the presence of DMSO 1 d after inoculation was defined as 1. The data shown here represent three independent experiments. At 14 d after inoculation, expression of HEV ORF2 capsid protein was determined via (C) western blot and (D) immunofluorescence microscopy. BF, bright-field microscopic image; FL, fluorescence microscopic image. P-values were determined by unpaired two-tailed Student’s t-tests (significant at *P < 0.05; **P < 0.01).