FIG 7.

Mutating the MAPV N protein to N:H386S or N:Δhvd fails to confer IFN regulation. (A and B) HEK293T cells were cotransfected with plasmids expressing an ANDV N-GFP fusion protein (ANDV-GFP) and plasmids expressing wt ANDV N protein, ANDV N:S386H, ANDV N:Δhvd, or ANDV N:Δhvd-S386H (A) or mutants with changes in the HVD and residue 386, ANDV N:Δhvd (ANDV N:Δhvd-S386H) and MAPV N:Δhvd (MAPV N:Δhvd-H386S) (B). Cell lysates were immunoprecipitated (IP) at 48 h posttransfection with anti-GFP antibody and assayed by Western blotting (WB) for coprecipitated ANDV N protein or input N protein by Western blotting. (C) HEK293T cells were cotransfected as described in the legend to Fig. 1 with plasmids expressing Flag-MDA5, ISRE firefly luciferase and Renilla luciferase reporters, and the indicated N protein mutants or empty vector. Luciferase activity was measured, Western blot analysis was performed, and the results were analyzed 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), as determined by one-way ANOVA with Tukey’s post hoc test.