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Fig. 6. Quantitative analysis of p65 translocation, I ${kappa}$ B ${alpha}$ degradation and NF- ${kappa}$ B-dependent transcription in single living cells. (A) Dual confocal microscopy images of p65-dsRed and I ${kappa}$ B ${alpha}$ -EGFP before and 40 minutes after TNF ${alpha}$ stimulation. (B) Quantitative analysis of the time course of p65-dsRed translocation, I ${kappa}$ B ${alpha}$ -EGFP degradation and cell luminescence (luciferase activity) in three cells, a-c (A,C). p65-dsRed translocation is shown as nucleo-cytoplasmic ratio and I ${kappa}$ B ${alpha}$ degradation is shown as EGFP fluorescence. Luminescence activity is shown as luminescence counts from successive luminescence images with background counts removed. (C) Luminescence images of cells following TNF ${alpha}$ stimulation. Luminescence directed by NF- ${kappa}$ B-promoter-directed luciferase activity. Cells marked a-c correspond to those marked in A and used for quantification in B.

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