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. 2014 May 22;8(5):e2891. doi: 10.1371/journal.pntd.0002891

Figure 3. Functional analysis of NS5 GTR sequence mutations in JEV infectious cDNA clone.

Figure 3

(A) Immunofluorescence analysis and plaque morphology of JEV genome-length viral RNA replication containing NS5 GTR sequence mutations in transfected BHK-21 cells at the indicated time points. Monoclonal antibody against SLEV envelope protein and Texas Red-conjugated goat anti-mouse IgG were used as primary and secondary antibodies, respectively. The plaque morphology of GTR sequence mutants and WT virus was determined by the double-layer plaque assay using the supernatants collected at 120 hpt. (B) Virus production of the transfected cells at each time point post transfection was detected by monolayer plaque assay, and the visible plaques were used to calculate titers of JEV WT and GTR mutants.