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Fig. 4. Inhibition of p65 nuclear translocation. (A) HeLa cells were incubated with medium containing the indicated peptide (30 µM) for 30 minutes, washed and then stimulated with TNF-R1-specific mutant TNF (100 ng/ml) for 30 minutes. Cells were then fixed and p65 was detected by immunofluorescence. (B) HeLa cells were incubated with increasing concentrations of Antp peptide for 30 minutes and then stimulated with the TNF-R1-specific mutant TNF (100 ng/ml). To quantify the inhibitory effect of Antp on p65 nuclear translocation, cells were subdivided into three groups and counted manually: cells with a clear nuclear staining (++), cells with equal levels of cytoplasmic and nuclear staining (+) and cells with fluorescence confined to the cytoplasm (–). For each condition more than 200 cells were counted and analysed in three independent experiments. Scale bars: 20 µm (A) and 50 µm (B).
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