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Fig. 5. Influence of bafilomycin A1 on the cellular localization of CPPs and on the nuclear translocation of p65. (A) HeLa cells were treated with Baf A1 (300 nM) for 30 minutes, washed and incubated with Antp or R9 (each 30 µM) for a further 30 minutes at 37°C before being examined by confocal microscopy at RT. (B) In a second experiment, after incubation with Baf A1 and peptide as above, cells were washed with medium and stimulated with the TNF-R1-specific mutant TNF (100 ng/ml) for 30 minutes. After fixation the distribution of p65 was determined by immunofluorescence microscopy. The change of the subcellular distribution of the CPPs upon fixation precluded the parallel detection of CPP localization and NF-
B translocation. Scale bars: 20 µm.
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