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. 2013 Dec 2;13(23-24):3442–3456. doi: 10.1002/pmic.201300001

Figure 2.

Figure 2

Inhibitory effects of JEV NS5 protein on IFNβ‐induced responses. (A) To analyze ISRE promoter activity, vector control and NS5‐expressing cells were transiently cotransfected with reporter plasmid containing firefly luciferase under control of ISRE and internal control reporter pRluc‐C1. After 4‐h IFNβ treatment, firefly luciferase and renilla luciferase were measured; firefly luciferase activity normalized to renilla luciferase activity is reported. (B) To analyze ISRE‐driven gene expression, vector control and NS5‐expressing cells were treated with or without 1000 U/mL IFNβ for 8 h; relative levels of PKR and OAS mRNAs were gauged by quantitative real‐time PCR, relative fold levels of PKR or OAS mRNA presented as ratio of PKR or OAS mRNA/GAPDH mRNA. (C) To analyze STAT1 phosphorylation, Western blot of lysates from cells treated with IFNβ for 0, 30, 60, or 120 min was performed by anti‐phospho‐STAT1 (Tyr701) and anti‐β actin antibody as internal control.