FIG 8.

The HA-NSs protein and Flag-MAVS bind in HEK 293T cells. HEK293T cells were cotransfected with Flag-MAVS, Flag-TBK1, and HA-EV or HA-NSs plasmids. At 24 h p.t., cells were lysed, and immunoprecipitation (IP) assays were performed using protein A/G agarose coated with the corresponding antibody. The beads were washed extensively 4 times and incubated with loading buffer at 95°C to rescue the supernatant, which was used for Western blotting. (A) Whole-cell lysate (WCL) of each sample and (B) IP fractions with the corresponding primary antibody highlighted on the left side of the panels. Western blotting was performed using mouse anti-TBK1, mouse anti-MAVS, mouse anti-HA, and mouse anti-GAPDH. horseradish peroxidase (HRP)-conjugated protein A/G was used to detect the primary antibodies.