FIG 12

GRP78 is an important host factor for JEV replication postentry. (A) Neuro2a cells were transfected with 300 ng JEV RNA, and at 6 h posttransfection, the cells were fixed and stained with JEV E antibody. (Left) DIC image. (Middle) JEV E staining. (Right) Merge of the two. Bar, 10 μm. (B to D) Neuro2a cells were transfected with NT or GRP78 siRNAs and 48 h later were transfected with 300 ng purified JEV RNA. At 24 h post-JEV RNA transfection, cells were processed for estimation of relative JEV RNA levels (B) and Western blotted for the indicated antibodies (D), and virus titers in the culture supernatant were determined by plaque assays (C). Each experiment contained biological duplicates, and the data presented in panels B and C are from three independent experiments. (D) The ratios of band intensities for different JEV antibodies and GAPDH are shown below the Western blots. The levels of viral RNA and titers in GRP78 siRNA-treated cells were compared to that in the NT siRNA-treated cells (**, P < 0.01). The error bars indicate SD.