FIG 2.

ANDV infection impairs MAVS-induced IRF3 activation. Huh-7 cells were mock infected (A) or infected with ANDV at an MOI of 1 (B and C). At 24 h p.i., cells were transfected with GFP-IRF3 together with a plasmid encoding FLAG-MAVS (A to C). At 24 h p.t., the cells were fixed using 4% PFA and permeabilized with PBS-Triton, and covers were incubated with a mouse anti-ANDV-N antibody (A and B). Alexa Fluor anti-mouse 594 antibodies were used as a secondary antibody, and Vectashield with DAPI was used as the mounting medium. (C) Covers were incubated with a mouse anti-FLAG together with a rabbit anti-ANDV-N antibody. Alexa Fluor anti-mouse 594 antibodies and Alexa Fluor anti-rabbit 350 antibodies were used as a secondary antibody, while Dako without DAPI was used as the mounting medium. The images were obtained by Olympus epifluorescence microscopy and processed with ImageJ. (D) The numbers of cells with a nuclear and cytoplasmic distribution of GFP-IRF3 from panels A (25 cells) and B (75 cells) were counted and results presented as a percentage. From panel B, only the GFP-IRF3 and ANDV-N protein-positive cells were counted. As a statistical test, a one-way ANOVA using the standard error of the mean (SEM) was performed.