FIG 6.

ANDV N protein S386 is critical for IFN regulation. (A and B) HEK293T cells were cotransfected as described in the legend to Fig. 1 with ISRE or IFNβ firefly luciferase reporters and plasmids expressing Flag-MDA5, control Renilla luciferase, and plasmids expressing the indicated N proteins: wt ANDV N, ANDV N:S386H, wt MAPV, or MAPV N:H386S. Luciferase activity was measured and Western blot analysis of input proteins was performed as described in the legend to Fig. 1. Assays were performed in triplicate with similar results in at least 3 separate experiments. Asterisks indicate statistical significance (*, P < 0.05; **, P < 0.01), as determined by one-way ANOVA with Tukey’s post hoc test. (C) HEK293T cells were cotransfected as described in the legend to Fig. 2 with plasmids expressing IRF3 or Flag-MDA5 and plasmids expressing the indicated N protein: wt ANDV N, ANDV N:S386H, MAPV N:H386S, or wt MAPV N. After 24 h, cells were harvested and analyzed by Western blotting as described in the legend to Fig. 2, and the results are representative of those from ≥2 experiments.