Overexpression of Hdj2 facilitates virus replication. A. HEK293T cells were mock-transfected or transfected with Flag-Hdj2 expressing plasmid. The cells were harvested at the indicated hour post-transfection (hpt), and subjected to Western blotting analysis using anti-Flag antibody for detection of Flag-Hdj2 expression. B. Transfected cells were infected with JEV. The total RNA and supernatant were harvested from mock or Hdj2-transfected at the indicated hour post-infection (hpi) and subjected to Northern analysis, or virus titer determination, respectively. The RNA was probed with a DIG-labeled oligonucleotide detecting nt 10950 to nt 10976 in the 3' UTR and an oligonucleotide probe detecting 18S rRNA. C. The amounts of RNA genome were normalized to that present in the 18S rRNA, and the relative amounts were plotted. D and E. The virus titers from each time point were calculated and plotted. F. JE virus titer was measured at 48 h post-infection of pEGFP-Hdj2, pEGFP, or mock-transfected cells. Error bars indicate the standard deviations of results from three independent experiments.