Fig. 1

Identifying the mutants. (A) The sequencing results of 10 passage cultures of the virus showed that the aa of the three mutants at E389 were different from those of the WT. (B) Statistical analysis of viral plaque diameters of the WT virus and mutants. Three independent experiments were conducted, with 10 plaques per virus were selected at random for measuring in each experiment. The data are presented as means and SD and were tested for significance using a one-way ANOVA with multiple comparison tests (**, P<0.01;***, P<0.001; ****, P<0.0001). (C) Plaque form and size of WT, along with three mutants, D389S, D389G, and D389G, in BHK-21 cells. (D) The BHK-21 cells were inoculated with viruses for 48 h, four mutants were identified using the JEV NS1 protein antibodies and stained with FITC-labeled antibodies. The identifiable cells exhibited green fluorescence.DAPI was used to stain the nucleus. The data are representative of at least three independent experiments