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. Author manuscript; available in PMC: 2019 Apr 19.
Published in final edited form as: Cell Rep. 2018 Jul 24;24(4):861–872.e6. doi: 10.1016/j.celrep.2018.06.045

Figure 2. batVP35 Inhibits IFN-β Production Independently of dsRNA Binding.

Figure 2

(A) In vitro dsRNA binding assay for eVP35 215–240 and batVP35 159–284. Fractional binding of batVP35 was normalized to eVP35, and error bars represent SD for the triplicate. RNA binding was assessed twice. (B) Western blot analysis of poly(I:C) pull-downs of the indicated FLAG-tagged VP35 constructs. IP, immunoprecipitation; WCL, whole-cell lysate. Poly(I:C) pull-downs were repeated twice. (C-E) IFN-β luciferase reporter assay stimulated by overexpression of (C) RIG-I N, (D) TBK1, or (E) IKKε in the presence of FLAG-tagged eVP35 or batVP35 (500 and 50 ng). Error bars represent the SEM for triplicate experiments. VP35 expression was assessed by western blot for the FLAG epitope tag, and the western blot was aligned to the corresponding samples in the graph. (F) IFN-β reporter assay in cells transfected as indicated. IFN-β promoter activity was assessed as in (C). VP35 expression was assessed for the highest concentration (500 ng) as in (C) (inset). Statistical significance was assessed using a oneway ANOVA and Tukey’s test: ****p < 0.0001, **p<0.01. E refers to empty vector. IFN-β luciferase reporter assays were repeated at least three times. See also Figures S2 and S3.