Figure 3

Effect of (+)sfRNA and (-)sfRNA on JEV RNA synthesis, when transfected into JEV-infected BHK-21 cells. A and B. Cells were either left uninfected (uninf., lane 1), or infected (lanes 2-10) with JEV at an MOI of 0.01. Plus-sense (+) (lanes 3, 6 and 9) and minus-sense (-) (lanes 4, 7 and 10) of sfRNAs were transfected at 28 hpi, or mock transfected (lanes 2, 5 and 8). Cytoplasmic RNA was extracted at the indicated hours post-infection (hpi). RNA was probed with a DIG-labeled minus-sense sfRNA to detect plus strands (A) or with plus-sense sfRNA to detect minus-strands (B). Oligonucleotide probe detecting 18S rRNA is shown at the bottom. C and D. Cells were either left uninfected (uninf., lane 1), or infected (lanes 2-7) with JEV at an MOI of 0.01. Minus-sense of sfRNAs (lanes 5-7) were transfected at 28 h postinfection (hpi), or mock transfected (lanes 2-4). Cytoplasmic RNA was extracted at the indicated time point. Dig-labeled riboprobes were used as indicated at the top. Oligonucleotide probe detecting 18S rRNA is shown at the bottom. Effect of transfecting (-)sfRNA into the JEV-infected BHK-21 cells on genome (E) or antigenome (F) was plotted. Error bars indicate the standard deviations of results from three independent experiments.