Journal: Journal of Virology
Article Title: Cellular DNAJA3, a Novel VP1-Interacting Protein, Inhibits Foot-and-Mouth Disease Virus Replication by Inducing Lysosomal Degradation of VP1 and Attenuating Its Antagonistic Role in the Beta Interferon Signaling Pathway
Figure Lengend Snippet: FMDV VP1 protein inhibits the SeV-induced IFN-ÃÂ² signaling pathway. (A) The expression of VP1 inhibits SeV-induced activation of IFN-ÃÂ² promoters. HEK293T cells were transfected with Flag-VP1 or empty vector, together with IFN-ÃÂ² luciferase reporter. At 24 hpt, the cells were mock infected or infected with SeV for 12Ã¢ÂÂh before luciferase assays were performed using a dual-specific luciferase assay kit. (B) VP1 negatively regulates SeV-induced activation of IFN-ÃÂ² at the IRF3 level. HEK293T cells were transfected with IFN-ÃÂ² reporter, pRL-TK, expression plasmids for Flag-VP1, and the indicated protein. Luciferase assays were performed with a dual-specific luciferase assay kit. Protein expression was analyzed by Western blotting. (C, D, and E) FMDV VP1 inhibits the phosphorylation, dimerization, and nuclear translocation of IRF3 after SeV stimulation. HEK293T cells were transfected with the indicated plasmids for 24Ã¢ÂÂh. Cells were infected with SeV at various time points and then harvested for analysis by Western blotting (C) or native-PAGE (D). (E) Cells were stained with the indicated antibodies and imaged by confocal microscopy.
Article Snippet: At 24 hpt, the cells were mock infected or infected with SeV for 12Ã¢ÂÂh; the luciferase activity was then measured by using a dual-specific luciferase assay kit (Promega).
Techniques: Expressing, Activation Assay, Transfection, Plasmid Preparation, Luciferase, Infection, Western Blot, Translocation Assay, Clear Native PAGE, Staining, Confocal Microscopy