Fig. 6. TRIP6 is involved in IL-1-, TLR2- and Nod1-induced NF-B activation. (A) TRIP6 potentiates IL-1-induced NF-B activation. 293 cells (1x10⁵) were transfected with 0.1 µg NF-B luciferase reporter plasmid and the indicated amounts of TRIP6 expression plasmid. 14 hours after transfection, cells were treated with IL-1 (10 ng ml^-1) (filled bars) or left untreated (empty bars) for 6 hours before luciferase assays were performed. (B) TRIP6 potentiates TLR2-induced NF-B activation. 293 cells (1x10⁵) were transfected with 0.1 µg NF-B luciferase reporter plasmid, 0.5 µg TLR2-encoding (filled bars) or empty control (empty bars) plasmid, and the indicated amounts of TRIP6 expression plasmid. Luciferase assays were performed 14 hours after transfection. (C) TRIP6 potentiates Nod1-induced NF-B activation. The experiments were done as in B except that TLR2 was replaced with 0.1 µg Nod1 expression plasmid. (D,E) A TRIP6 mutant inhibits IL-1-, TLR2- and Nod1-induced NF-B activation. 293 cells (1x10⁵) were transfected with 0.1 µg NF-B luciferase reporter plasmid and 0.5 µg TRIP6-(280-480)-encoding (filled bars) or empty control (empty bars) plasmid. 14 hours after transfection, cells were treated with IL-1 (10 ng ml^-1) or left untreated for 6 hours before luciferase assays were performed (D). Alternatively, 293 cells (1x10⁵) were transfected with 0.1 µg NF-B luciferase reporter plasmid, 0.5 µg TRIP6-(280-480)-encoding (filled bars) or empty control (empty bars) plasmid, and 0.5 µg of TLR2 (D) or Nod1 (E) expression plasmid. Luciferase assays were performed 14 hours after transfection. (F) TRIP6 dominant-negative mutant has no effect on IFN--induced IRF-1 activation. 293 cells (1x10⁵) were transfected with 0.1 µg IRF-1 luciferase reporter plasmid and 1 µg TRIP6-(280-476)-encoding (filled bars) or control (empty bars) plasmid. 14 hours after transfection, cells were treated with IFN- (100 ng ml^-1) or left untreated for 6 hours before luciferase assays were performed.