Equine Arteritis Virus Does Not Induce Interferon Production in Equine Endothelial Cells: Identification of Nonstructural Protein 1 as a Main Interferon Antagonist
Figure 6
Effect of EAV nsp1 in IRF-3 and NF-κB nuclear translocation. (a) HeLa cells were transfected with FLAG-tagged EAV nsp1 for 12 h and cells were stimulated with poly(I:C) (1 μg/mL) for 8 h. Cells were fixed and stained with rabbit anti-IRF-3 Ab for endogenous IRF-3 and mouse anti-FLAG Ab for FLAG-tagged EAV nsp1, followed by incubation with Alexa Fluor 594-conjugated (red) and 488-conjugated (green) secondary antibodies, respectively, along with DAPI for nucleus staining (blue). (b) HeLa cells were transfected with FLAG-tagged EAV nsp1 for 12 h and subsequently treated with 20 ng/mL of TNF-α for 45 min or untreated. Cells were then costained with mouse anti-FLAG MAb for FLAG-tagged EAV nsp1 and rabbit anti p65 polyclonal antibody for endogenous NF-κB p65 subunit followed by Alexa Fluor 488-labeled (green) anti-mouse Ab and Alexa Fluor 594-labeled (red) anti-rabbit Ab, respectively, along with DAPI for nucleus staining (blue).
