FIG 1.

MAPV N protein fails to regulate ISRE and IFNβ induction. HEK293T cells were cotransfected with a constant amount of total DNA using plasmids expressing ANDV, NY-1V, or MAPV N proteins or pcDNA3.1⁺, ISRE, or IFNβ promoter-directed firefly luciferase (Luc) reporters, an internal pRL-null Renilla luciferase control, and IFN pathway-activating plasmids expressing Flag-MDA5 (A to C), Flag-RIG-I-CARD (D), or Flag-TBK1 (E and F). Firefly luciferase activity was measured at 24 h posttransfection, normalized to control cotransfected constitutively expressing Renilla luciferase activity, and reported as the fold increase compared to that in the empty vector pcDNA3.1⁺-transfected controls. Western blot analysis of N protein, pathway inducers, and β-actin (total protein) indicates comparable protein expression levels in the lysates. The assays were performed in triplicate with similar results in at least 3 separate experiments. Asterisks indicate statistical significance (*, P < 0.05; **, P < 0.01; ***, P < 0.001), as determined by one-way ANOVA with Tukey’s post hoc test.