Fig. 1. Nuclear accumulation of p65-EGFP in response to TNF stimulation. (A) Time series images of p65-EGFP fluorescence at stated times (in minutes) after addition of TNF. Inhibition of translocation was observed in cells treated with 18 µM SN50 for 15 minutes prior to addition of TNF. (B) Quantification of p65-EGFP translocation in response to TNF. Mean nuclear and cytoplasmic fluorescent intensities were determined for fluorescent cells at each time point and plotted as a ratio relevant to the initial ratio at t=0 minutes. Translocation is shown in the absence of inhibitor or in the presence of 18 µM SN50, 18 µM SN50M or 12.5 mM Bay11-7082. Confocal microscopy was carried out in cells transfected with p65-EGFP 24 hours prior to their use. Cells were treated with appropriate inhibitor for 15 or 30 minutes (see text) prior to stimulation with 10 ng/ml TNF 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.