Figure 2.

Induction of NF-κB in KT-3 cells by CD30. (A) KT-3 cells were transiently transfected with pcDNA3-based vectors for expression of full-length (TRAF1 or TRAF2) and truncated (ΔTRAF1 or ΔTRAF2) proteins, as indicated. After overnight incubation, KT-3 cells were stimulated by anti-CD30 mAb (C10) for 1 hr, and nuclear extracts were prepared. EMSAs were performed as described. Similar results were obtained in three independent experiments. (B) Analysis of the NF-κB-binding complex induced by anti-CD30 mAb. Unlabeled oligonucleotide competitors were used at 100-fold molar excess. For supershift analysis, nuclear extracts from KT-3 cells were preincubated with antiserum to p50, p65, and normal rabbit serum (NRS). Similar results were obtained in three independent experiments. The open arrow denotes the NF-κB complexes.