Fig 7.

WEEV infection inhibits neuronal PRR pathway activity. (A) BE(2)-C/m ISRE promoter-reporter cells were infected with WEEV at an MOI of 1 for 1.5 h and subsequently treated with the indicated stimulus (left graph) or pretreated with the indicated stimulus for 3 h prior to infection with WEEV (right graph), and both reporter activity and viability were assessed relative to mock-infected controls at 16 to 20 hpi. We used 100 U/ml IFN-α/β-A/D, 50 μg/ml extracellular poly(I-C) (pIC), and 500 ng/ml transfected poly(I-C) (T-pIC) for stimulation. *, P ≤ 0.05 compared to appropriate mock-infected control. (B) BE(2)-C/m NF-κB promoter-reporter cells were treated and analyzed as described above for panel A except that 25 ng/ml TNF-α was used instead of IFN-α/β-A/D. (C) BE(2)-C/m cells were mock infected (upper gel in each gene-specific group) or infected with WEEV at an MOI of 1 for 3 h (lower gel in each gene-specific group) and treated with the indicated stimulus, and transcripts for the indicated genes were analyzed by semiquantitative RT-PCR 4 h after stimulation. Adjacent lanes for individual samples represent results using 10-fold dilutions of cDNA. (D) BE(2)-C/m cells were infected and stimulated as described in panel B except that WEEV was used at an MOI of 10 and gene transcripts were analyzed by qRT-PCR 4 h after stimulation. **, P < 0.0001.