Supplementary Figure 5: RNF128 positively regulates TBK1 kinase activity.

(a) TBK1kinase activity assays in HEK293 cells transfected with the RNF128 expression plasmid and then infected with SeV for 6 h. (b) TBK1 kinase activity assays in peritoneal macrophages transfected with mice RNF128 siRNA, then the cells were infected with SeV for 6 h. (c) Quantitative RT-PCR analysis of mRNA expression of Ifnb1 (upper) and western blot analysis of phosphorylated-TBK1 protein (lower) in TBK1 deficient MEFs transfected with expression plasmids for Myc-TBK1-WT or Myc-TBK1-S172A along with V5-RNF128 plasmids. *p <0.05, **p <0.01, ns, not significant (two-tail Student's t-test). Data are from three independent experiments (a,b; mean±S.D. of triplicate assays) or are representative of three independent experiments with similar results (c-h).