NOG1 did not affect the phosphorylation and nuclear translocation of IRF3.

(A) WT and NOG1+/- cells were mock-infected or infected with SeV for 12 h, the cells were collected, and the levels of IRF3 phosphorylation were detected by Western blotting. (B) HEK-293T cells were transfected with 3 μg of Flag-NOG1-expressing plasmid. At 24 hpt, the cells were mock-infected or infected with SeV for 12 h. Cells were harvested and subjected to nuclear and cytoplasmic fractionation. The expression of IRF3 in the cytoplasm (Cyt) and nucleus (Nuc) was detected by Western blotting, respectively. Histone 3.1 was used as a nuclear loading control and marker, and β-actin was used as a cytosolic loading control and marker. The abundance of IRF3 was quantified using ImageJ software. (C) HEK-293T cells transfected with 0.5 μg of Flag empty vector or Flag-NOG1-expressing plasmid were mock-infected or infected with SeV for 12 h. The subcellular localization of NOG1 and IRF3 was detected by IFA. (TIF)