AMS inhibited JEV infection in BHK21 cells. (A) AMS cytotoxicity was assessed by the viability of BHK21 cells treated with AMS. Pre-seeded BHK21 cells were treated with increasing concentrations of AMS for 24 h, and then subjected to cell viability analysis using CCK-8. (B,C) Plaque reduction assay to determine the effect of AMS on JEV infection in BHK21 cells. BHK21 cells were treated with indicated concentrations of AMS during the addition of JEV strain SA-14-14-2 (100 pfu) and the inhibition efficiency was measured with plaque assay (B). Plaques were counted and percentage of plaque reduction was calculated (C). (D,E) AMS affected JEV strain SA-14-14-2 replication in BHK21 cells in a dose-dependent manner. JEV strain SA-14-14-2 was incubated with indicated concentrations of AMS and infected BHK21 cells at an MOI of 0.1. At 48 h p.i., the intracellular level of JEV E protein (green) in BHK21 cells was detected with immunofluorescence analysis (D). The titer of JEV in the supernatant was quantified by plaque assay (E). Bars, 50 μm. Data are means ± SD of triplicate experiments.