FIG. 5.

Reconstitution of TRAIL-induced IKK and JNK activation in RIP^−/− and TRAF2^−/− cells. (A) Wild-type, RIP^−/−, and TRAF2^−/− mouse fibroblasts cells were cotransfected with HA-IKKβ and Myc-RIP or Flag-TRAF2 expression plasmids as indicated. Twenty four hours after transfection, cells were treated with GST-TRAIL (10 μg/ml) for 30 min and subjected to an IKK assay. HA-IKKβ, Myc-RIP, and Flag-TRAF2 were detected by Western blotting. Nontreated cells served as controls. (B) Wild-type, RIP^−/−, and TRAF2^−/− mouse fibroblasts were cotransfected with HA-JNK1 and Myc-RIP or Flag-TRAF2 expression plasmids as indicated. Cells were treated with GST-TRAIL (10 μg/ml) for 30 min and analyzed by JNK assay. HA-JNK1, Myc-RIP, and Flag-TRAF2 were detected by Western blotting. Nontreated cells served as controls.