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Fig. 2. TRIP6 is involved in RIP2-mediated NF-B activation. (A) TRIP6 weakly activates NF-B in a dose-dependent manner. 293 cells (1x10⁵) were transfected with 0.1 µg NF-B luciferase reporter plasmid and the indicated amounts of HA-tagged TRIP6 expression plasmid. Luciferase assays were performed 14 hours after transfection. TRIP6 expression levels in the transfected cells were analysed by western blot with anti-HA antibody (bottom). (B) TRIP6 synergizes with RIP to activate NF-B. 293 cells (1x10⁵) were transfected with 0.1 µg NF-B luciferase reporter plasmid, 0.5 µg Flag-RIP2 expression plasmid and the indicated amounts of HA-TRIP6 expression plasmid. Luciferase assays were performed 14 hours after transfection. Increased TRIP6 expression did not affect RIP2 levels as suggested by western blots with anti-Flag and anti-HA antibodies (bottom). (C) Effects of TRIP6 deletion mutants on RIP2-mediated NF-B activation. 293 cells (1x10⁵) were transfected with 0.1 µg NF-B luciferase reporter plasmid, 0.5 µg Flag-RIP2 expression plasmid (empty bars) or empty control plasmid (filled bars), and 0.5 µg of plasmids encoding the indicated HA-tagged TRIP6 mutant. Luciferase assays were performed 14 hours after transfection. Expression levels of RIP2 and the TRIP6 mutants were analysed by western blots with anti-Flag and anti-HA antibodies respectively (bottom).

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