Figure 3. Nedd4 does not affect JEV entry.

SK-N-SH cells were transfected with different concentrations of Nedd4-specific siRNA or the NT siRNA (50 nM) for 72 h to establish gene knockdown. (A) SK-N-SH cells were incubated with JEV (MOI = 10) at 4 °C for 1 h and then collected. The binding of JEV RNA was detected by RT-qPCR. (B) SK-N-SH cells were incubated with JEV (MOI = 10) at 4 °C for 1 h, and the cells were then transferred to 37 °C for another 1 h. A JEV internalization assay was performed, and JEV RNA was analysed by RT-qPCR. (C) SK-N-SH cells were infected with JEVpv, and the infectivity was assessed by identifying GFP-positive cells. The nuclei were stained with DAPI (blue). Images were obtained using a confocal microscope (left panel, scale bar = 50 μm) and quantitated using ImageJ software (right panel). The data are shown as the mean ± SD of three independent experiments.