MAVS activates TBK1 and IKKε through TRAFs in NEMO dependent and independent manner
Fig 4
TRAFs’ E3 ligase activity is required for TBK1/IKKε activation via NEMO.
(A) and (B) WT 293T cells, 293T cells with combined deficiency of TRAFs and NEMO (TRAFs/NEMO-deficient) and the same deficient cells reconstituted with TRAF2, 3, or 6 or their dRing deletions were infected with SeV for the indicated times. Cells were analyzed by Western blot to detect the phosphorylation and expression of the indicated proteins (A), or RT-PCR analysis of the indicated gene induction (B). (C) WT 293T cells, TRAFs-deficient and TRAFs/NEMO-deficient cells were transiently transfected with TRAF2, 3, or 6 or their dRing deletions for 12 h and infected with SeV for the indicated times. Cells were analyzed by Western blot to detect the phosphorylation and expression of the indicated proteins. (D) Cells in the left panel of (C) were analyzed by ELISA to detect the production of IL6. Data from (D) represent mean ± SD. Similar results were obtained in 3 independent experiments. See also S5 Fig.