Comparison of the effects of ASA and SSA on NF-κB activation in Jurkat cells.
(A) Cells were treated with ASA (2 × 10−3 M), SSA (2 × 10−3 M), or corresponding amounts of DMSO carrier before stimulation with A23187 (0.5 μg/mL) with or without PMA (10 ng/mL) for 2 hours. Nuclear extracts from each sample were incubated with a radiolabeled consensus NF-κB oligonucleotide. The arrows indicate 2 complexes induced by stimulation with A23187 and PMA (lane 2) but not A23187 alone (lane 5). Both complexes contained NF-κB molecular species as detected by coincubation with anti-p65 (lane 3) or anti-p50 antibodies (lane 4). (B) Jurkat cells transiently transfected with the IL-4.265 plasmid were treated (15 minutes) with ASA (2 × 10−3 M), SSA (2 × 10−3 M), 5-ASA (2 × 10−3 M), or corresponding amounts of DMSO before 20-hour stimulation with A23187 (0.5 μg/mL). The mean ± SEM percentage of control CAT expression in 4 independent experiments done in duplicate is shown. The asterisk indicates P < .05 (Wilcoxon test) relative to samples treated with DMSO.
