FIG 5.

JEV NS1′ inhibits the miR-22-mediated IFN-β response via CDK1. (A) HeLa cells were transfected with pcDNA3.1 plasmids encoding the JEV NS1′ protein or an empty vector. After 24 h transfection, the cells were treated with 5 μg/ml RO3306 or DMSO for a period of 12 h. The mRNA level of miR-22 was analyzed by qRT-PCR. (B) HeLa cells were subjected to transfection and RO3306 treatment paradigm as described in panel A, followed by poly(I·C) treatment for 12 h. The mRNA level of IFN-β was analyzed by qRT-PCR. (C and D) HeLa cells treated with 5 μg/ml RO3306 or DMSO for 2 h were infected with the WT virus or NS1′-def virus at an MOI of 1.0. At 24 hpi, the mRNA level of IFN-β was determined by qRT-PCR (C) and the viral titer in the supernatant was measured by plaque assay (D). All data were pooled from three independent experiments. **, P < 0.01; ***, P < 0.001. ns, nonsignificant.