Figure 5.

TRAM activates NF-κB and is specific to the TLR4 pathway. (a) HEK293 cells were transfected with 40 ng of an NF-κB reporter construct and cotransfected with TRAM, TRAM-TIR, TRAM-C113H, and TRAM-P112H. (b) TLR-expressing HEK293 stable cell lines were transfected with 40 ng of an NF-κB reporter gene and cotransfected with increasing concentrations of TRAM-C113H. 1 d after transfection, TLR-expressing cells were stimulated with Malp-2 (2 nM), dsRNA (100 μg/ml poly IC), LPS (10 ng/ml), R-848 (10 μM), IL1β (10 ng/ml) or TNFα (10 ng/ml) or left untreated or 8 h, and luciferase reporter gene activity was measured.