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. 2020 Apr 15;11:656. doi: 10.3389/fmicb.2020.00656

FIGURE 4.

FIGURE 4

HEV ORF2 protein inhibits TRIF and MyD88 induced IFN-β promoter activation. (A) TRIF assay was performed by co-transfecting 0.5 ng myc-TRIF plasmid with two different quantities of FL ORF2 (10 and 25 ng) along with IFN-β firefly and TK Renilla luciferase reporter plasmids. Luciferase activity was measured 24 h post-induction. (B) MyD88 assay was performed by co-transfecting 2.5 ng of MyD88 plasmid with two different quantities of FL ORF2 (10 and 25 ng) along with IFN-β firefly and TK Renilla luciferase reporter plasmids. Luciferase activity was measured 24 h post-induction. (C) Cells were co-transfected with TRIF (20 ng) and FL ORF2 plasmids (1 μg each). RNA was isolated 24 h post transfection. Ct values corresponding to IFN-β and (D) The ISG56 mRNA levels were normalized to that of the GAPDH Ct values and change in the mRNA level was calculated by the ΔΔCt method with respect to the un-induced vector control. All experiments were performed in HEK293T cells. Values are percent mean ± SD, n = 3 for all experiments (∗∗∗ denotes p-values ≤ 0.001).