Duck Enteritis Virus VP16 Antagonizes IFN-β-Mediated Antiviral Innate Immunity

Figure 2

DEV VP16 is having a demonstrable effect on IFN-β activation and IFN-β-related ISGs. (a) Duck IFN-β mRNA relative levels in DEF transfected 4 μg or 10 μg pCAGGS-UL48-HA, stimulated with 25 μg/mL poly (I:C) for over 36 hours at 12 hours posttransfection, were presented by real-time qPCR. The expression of VP16 was analyzed by western blotting using anti-HA and anti-β-actin (as a control) (CST, America). In this group, an increased amount of VP16-HA expression plasmid was used. (b) DEFs seeded in 24-well plates were transfected with pCAGGS or pCAGGS-UL48-HA (500 ng/well), together with duIFN-β-Luc (500 ng/well) and pRL-TK (50 ng/well) for 12 hours; then, 25 μg/mL poly (I:C) was transfected into cells for another 24 hours. Firefly luciferase activities were measured and the data were analyzed by GraphPad Prism software, and results were presented using two-way ANOVA (). (c, d) Quantitative real-time PCR analysis of duck OASL and duck Mx mRNA levels in DEF cells which express VP16-HA or not. Duck ISG mRNA relative expression was upregulated by (c) poly (I:C) or (d) poly (dA:dT). Experiments were shown using two-way ANOVA (). Significant, , , and . (e) Expression of VP16-HA was confirmed by western blot analysis.

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