Fig 3. Viperin and MAVS interact.

(A) 293T cells were untransfected (with only the transfection reagent present), or transfected with combinations of empty vector, pcDNA3.1-viperin, or pCMV-FLAG-MAVS. Cell lysates were subjected to immunoprecipitation (IP) with anti-viperin beads (V) or anti-MAVS beads (M) and blotted for viperin or MAVS. (B) RAW cells were stimulated with IFN (10³ units/ml) or transfected with poly(I:C) (1μg/ml) for 8h. Cell lysates were subjected to immunoprecipitation and blotted for viperin, MAVS, or calnexin as a negative control. (C) 293T cells were co-transfected with pcDNA3.1-viperin and pCMV-FLAG-MAVS and solubilized in 1% CHAPS in PBS. Equal aliquots were immunoprecipitated with anti-viperin beads or anti-MAVS beads. Beads were collected (V1 and M1 respectively), and the supernatants added to fresh beads for second (V2, M2) and third (V3, M3) immunoprecipitation. After immunoprecipitation, beads were collected (V2 and M2), and the process repeated. WCL is whole cell lysate without immunoprecipitation and Vsup and Msup designate the residual lysates after the third round of precipitation.