Fig 4.
Induction of IFN-β by EAV RNA in MEFs is MAVS dependent, and the recognition of EAV RNA primarily relies on the PRR MDA5. To investigate the role of RLR sensors in the recognition of arteriviral RNA, MEFs lacking MAVS, MDA5, or RIG-I (black bars) and their respective wild-type controls (white bars) were transfected with total RNA isolated from Vero cells infected with EAV (A), mengovirus (B), or NDV (C). At 3 h p.t., total RNA was isolated from these cells and used as a template for cDNA synthesis. Gene expression levels of IFN-β and GAPDH were subsequently determined by means of qRT-PCR. Using the comparative CT method, the gene expression levels of IFN-β were normalized to those of GAPDH, which was used as an internal standard, and compared to the gene expression levels of IFN-β in MEFs transfected with total RNA from mock-infected Vero cells. This experiment was performed three times independently with similar outcomes. Bars represent the mean of PCR triplicates from one representative experiment ± standard deviation.