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Fig. 3. Modulation of p65-dsRed translocation by glucocorticoids. HeLa cells expressing p65-dsRed and either EGFP (A) or EGFP-GR (B) were pretreated for 40 minutes with medium, 10 nM dexamethasone (Dex) or 10 nM RU486, followed by confocal microscopy (data not shown). Cells were then treated with 10 ng ml-1 TNF{alpha} and monitored every 2 minutes by confocal microscopy. p65-dsRed subcellular fluorescence was analysed and plotted as a ratio of nuclear:cytoplasmic fluorescence relative to the ratio at 0 minutes. (C) The time of 50% maximal nuclear import to 50% nuclear export of p65-dsRed from the data represented in A and B was determined for each treatment and plotted as the half-life of nuclear occupancy. Calculation of the 5% least significance difference between treatments showed all to be significantly different to one another (*) with the exception of EGFP and EGFP-GR transfected cells pretreated with dexamethasone. Each treatment was performed in triplicate with a minimum of 20 cells per experiment. Data plotted represent mean ratio per cell ± s.d.(n–1) for each experiment.