Fig. 4.
RSV NS2 inhibits the interaction between RLRs and MAVS. (A) The coIP between FLAG-RIG-IFL or FLAG-MDA5FL with HA-tagged MAVS and increasing concentrations of HA-NS2 (1, 2, and 4 μg) in the absence (Top) or presence (Bottom) of poly I:C. WCL, whole-cell lysate. (B) In vitro pull-down assays between NS2 and RLR CARDs. Recombinantly expressed and purified MBP-RIG-ICARDs (Top Left), MBP-MDA5CARDs (Top Right), or MBP tag alone (Bottom Left) were immobilized on amylose resin prior to incubation with NS2. NS2 alone was also incubated with amylose resin (Bottom Right) as a negative control. Lanes correspond to the following: M, marker; 1, bound beads; 2, input; 3, wash 1; 4, wash 4; 5, final beads; 6, elution. Shown are representative results repeated in triplicate.