Figure 4
Functional domain mapping of RNF166.
(A) Schematic structures of RNF166 and the deletion mutants used in this work. (B,C) Effects of truncations of RNF166 on SeV-induced (B) and VISA-mediated (C) activation of the IFN-β promoter. Transfection and luciferase assays were performed as above. (D) Effects of RNF166 and the RNF166 RING deletion mutant on VISA-mediated antiviral activity. 293T cells (5 × 105) were transfected with the indicated plasmids (1 μg). At 20 h after transfection, cells were infected with NDV-eGFP at an MOI of 0.0001 and at 40 h after infection, viral replication was determined by fluorescence microscopy. The GFP-positive cells were counted and the percentage was calculated. (E) Effects of RNF166 and the RNF166 RING deletion mutant on replication of SeV. 293T cells (5 × 105) were transfected with the indicated plasmids (1 μg). At 12 h after transfection, cells were infected with SeV and at 24 h after infection, mRNA of P protein was determined by Q-PCR. (F) RNF166 interacted with TRAF3 and TRAF6 via its zinc-finger domain. Transfection and immunoprecipitation (IP) were performed as in Fig. 2 (D). Each graph represents the mean ± SD of three independent experiments done in triplicate. ***indicates P < 0.001; **indicates P < 0.01; ns (not significant) indicates P > 0.05.
