Fig 5. Viperin suppresses MAVS-dependent IFNβ induction.

(A) WT and viperin KO BMM were treated with IFN at 10³ units/ml, or transfected with poly(I:C) at 1μg/ml or 5’ppp-dsRNA at 0.1μg/ml for 8 hours. qPCR analysis was performed to quantitate viperin and IFNβ mRNA expression. Results are presented as ±SEM of 4 independent experiments. (B) 293T cells were left untransfected or transfected with various combinations of the IFN reporter construct, MAVS, and viperin, and empty vector pcDNA3.1-myc-his where necessary to ensure equal amounts of DNA. Transfected cells were left untreated or transfected with poly(I:C) for 8 hours then subjected to luciferase activity assays. Results are presented as ±SD of quadruplicates. (C) 293T cells were transfected with 0.8μg of varying combinations of pCMV-FLAG-MAVS, pcDNA3.1-viperin, or, as a control, pcDNA3.1-Venus. Cells were lysed and lysates subjected to luciferase activity assays. Results are presented as ±SD of quadruplicates. (D) 293T cells were transfected with the IFN reporter construct and MDA-5 alone or together with viperin and subjected to luciferase activity assays. Results are presented as ±SD of quadruplicates.