Journal: Journal of Virology
Article Title: Blocking of Interferon-Induced Jak-Stat Signaling by Japanese Encephalitis Virus NS5 through a Protein Tyrosine Phosphatase-Mediated Mechanism
Figure Lengend Snippet: Inhibition of IFN-stimulated nuclear translocation of Stat1-DsRed by JEV NS5 in BHK-21 cells. BHK-21 cells were cotransfected with plasmids expressing the indicated JEV proteins and pStat1-DsRed. Twenty-four hours after transfection, cells were stimulated with or without IFN-αA/D (2,500 U/ml) for 16 h. Cells were then fixed and permeabilized for immunofluorescence assay. JEV proteins (C, prM, E, NS1, NS2A, NS2B, NS2B3, NS4A, NS4B, and NS5) were detected using a mouse anti-Flag antibody and Alexa-Fluor 488 goat anti-mouse antibody (green). JEV prM-E and NS3 were detected using anti-JEV E and NS3 antibodies, respectively, and then stained with Alexa-Fluor 488 goat anti-mouse antibody (green). Nuclear translocation of Stat1-DsRed (red) was observed under a fluorescence microscope. Representative cells from the same field for each experimental group are shown.
Article Snippet: The cells were probed with anti-Flag M2 antibody and subsequently with Alexa-Fluor 488 goat anti-mouse immunoglobulin G (Molecular Probes).
Techniques: Inhibition, Translocation Assay, Expressing, Transfection, Immunofluorescence, Staining, Fluorescence, Microscopy