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Fig. 2. Degradation of I{kappa}B{alpha}-EGFP in response to TNF{alpha} stimulation. (A) Time series images of I{kappa}B{alpha}-EGFP fluorescence at stated times (in minutes) after addition of TNF{alpha}. The degradation was prevented in cells treated with 12.5 µM Bay11-7082 for 30 minutes prior to addition of TNF{alpha}. (B) Degradation of I{kappa}B{alpha}-EGFP in response to TNF{alpha}. Mean cellular fluorescence intensities were determined for fluorescent cells at each time point and plotted as a percentage of the fluorescence values at t=0 minutes. Confocal microscopy was carried out in cells transfected with I{kappa}B{alpha}-EGFP 24 hours prior to their use. Cells were treated with appropriate inhibitor for 15 minutes (for SN50 and SN50M) or 30 minutes (for Bay11-7082) prior to stimulation with 10 ng/ml TNF{alpha} at t=0 minutes. Results show mean values±s.d.(n-1) from two experiments with at least six cells per experiment plotted for each treatment.