Figure 1. WT but not Optn^470T expression inhibits activation of NF-κB and IRF3.

Human optineurin constructs, homologous to murine WT (1–584), the Ub-binding point-mutant D477N, and the C-terminal truncation (1–470) are depicted (A). Human embryonic kidney 293 cells were transiently transfected with the indicated constructs and stimulated or not with TNF. Reporter luciferase activity was normalized to β-galactosidase activity (B, left panel), and anti-Flag blotting for Flag-optineurin variants is shown (B, right panel). hTLR3-293 cells, which stably express TLR3, were transiently transfected as indicated and stimulated or not with poly I:C. Luciferase activity (C, left panel), and anti-Flag blotting (C, right panel) were performed as in panel B. An average of duplicate samples of one representative experiment out of three is shown.