Figure 4

Detection of viral NS3 protein in NTZ-treated cells. BHK-21 cells were infected with JEV at a MOI of 0.001 and treated with NTZ at 3 μg/ml. The cells were harvested at 24 or 60 h post-infection (hpi). (A) The same amount (20 μg) of cell lysate was loaded in each lane. The expression of viral NS3 protein was examined by western blot. (B) The viral NS3 protein (green fluorescence) was detected at 24 hpi by immunofluorescence analysis. The cells were also stained for DNA with 4′, 6′-diamidino-2-phenylindole (DAPI, blue fluorescence). The Merge panels show the superimposed images. (C) Fluorescence intensity of NS3 and DAPI was quantified using the software ImageJ. The ratio of fluorescence intensity between NS3 and DAPI was calculated and plotted. The data are means with standard errors from three independent experiments. *, p < 0.01 between the groups tested.