MAVS50 targets TRAF molecules to inhibit MAVS70-mediated IFN induction.

<p>(A) Diagram of key signaling molecules of the RIG-I-dependent IFN induction. (B and C) 293T cells were transfected with an IFN-β reporter cocktail, a plasmid containing RIG-I-N (B) or TBK-1 (C), and increasing amount of plasmid containing MAVS50. The IFN-β promoter activity was determined by luciferase assay at 30 hours post-transfection. <i>**p<0</i>.<i>01</i>; <i>***p<0</i>.<i>005</i>. (D) 293T cells were transfected with plasmids containing MAVS wild-type (WT) or MAVS50 and a plasmid containing TRAF6. Whole cell lysates (WCL) were precipitated with anti-Flag (TRAF6). Precipitated proteins and WCLs were analyzed by immunoblotting with indicated antibodies. (E) 293T cells, depleted with endogenous MAVS and “reconstituted” with MAVS70 or MAVS50, were precipitated with anti-V5 (MAVS70 or MAVS50). Precipitated proteins and WCLs were analyzed by immunoblotting with indicated antibodies. (F) MAVS knockdown cells “reconstituted” with MAVS70-V5 were transfected with increasing amount of MAVS50-Flag. At 30 hours post-transfection, cells were infected with Sendai virus (SeV, 200 HA unit/ml) for two hours. WCLs were prepared and precipitated with anti-V5 (MAVS70) antibody. Precipitated proteins and WCLs were analyzed by immunoblotting with indicated antibodies. V, vector.