Figure 5.

NSs interact with Jak1 independently and inhibits its phosphorylation. (A) HEK293 cells expressing NT sgRNA or sgRNA for STAT1 were cotransfected with plasmids expressing V5-Jak1 and FLAG-tagged NSs. IP was performed 24 h post-transfection using an anti-V5 antibody. Total cell lysates (TCL) and IP products were analyzed by western blot using the indicated antibodies. (B) SFNV and SFSV infection inhibits Jak1 phosphorylation. A549 cells were infected with SFNV or SFSV. The next day the cells were treated with IFNα (1000 U\ml) for 30 mins. Western blot analysis was performed as described above using Jak1 and phospho-Jak1 specific antibodies. Tubulin was used as a loading control. Numbers below the bands represent average bands intensity from three independent experiments quantified using ImageJ and are expressed as a percent of mock-infected and stimulated control. (C) SFNV and SFSV infection inhibits IFNλ2 signaling. A549 cells were infected with SFNV or SFSV. The next day the cells were stimulated with IFNλ2 (70 ng\ml) for 20 mins. Western blot analysis was performed using total STAT1 and phospho- STAT1specific antibodies. Tubulin was used as a loading control.