Activation of NFκB by TRAIL. (A) Jurkat cells were incubated with PMA (50 ng/mL)/Ionomycin (2 μg/mL) (P/I) or TRAIL (0.3 μg/mL) (TR) for the time periods indicated. Cells were harvested, cellular proteins were isolated, and EMSA was performed. sc, specific competition with unlabeled oligonucleotide. Similar results were obtained in three independent experiments. (B) Jurkat cells were stimulated with TRAIL (0.3 μg/mL) for 30 minutes in the presence or absence of LLnL (6.25 μmol/L, 1 hour of pretreatment). Fifteen micrograms of protein extract was run on a 12% polyacrylamide gel. Proteins were transferred onto a nylon filter that was subsequentially hybridized with an anti-IκBα antibody. Similar results were obtained in three independent experiments. Similar results were obtained with CEM cells.