Fig. 7.

DDX5 interacts with the JEV core, NS3, NS5 proteins and colocalizes with them during virus infection. (A). DDX5 can interact with JEV core, NS3, NS5 proteins. BHK-21 cells was infected with JEV (MOI = 0.01). 48 h later, the anti-DDX5 polyclonal antibody was used to co-immunoprecipitate the proteins from the whole cell lysate treated with RNase (100 μg/ml) and DNase (100 μg/ml). The JEV core, NS3 and NS5 proteins were detected using their polyclonal antibodies respectively. The cell lysates (input, 2%) were analyzed in parallel by western blot. (B). DDX5 colocalizes with the JEV core, NS3, NS5 proteins. We infected the BHK-21 cells with JEV (MOI = 0.01) first. 48 h later, DDX5, JEV core, NS3 and NS5 proteins were detected using their polyclonal antibodies and the colocalization study was performed under confocal microscopy, the negative serum (the serum of unimmunized mouse) was used as the control. (C). DDX5 is not incorporated into the virions. BHK-21 cells were infected with JEV (MOI = 0.01) for 48 h, the anti-E monoclonal antibody was used to immunoprecipitate the mature virions in the medium, the proteins in the immunoprecipitated virions (IP) were detected with anti-DDX5, E or core antibodies. All cell lysates (input, 2%) were analyzed in parallel by western blot. The heavy chain of anti-E monoclonal antibody was labeled.