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Fig. 2. The UVB-induced second peak of PARP activation. (A) Time course of oxidant formation in UVB-irradiated cells. Cells exposed to 1.6 kJ/m² UVB were loaded with DCF-DA prior to harvesting to detect the oxidant load in the cytoplasm by immunofluorescence. (B) The quantification of oxidant signal in UVB-exposed cells. The cytoplasmic fluorescence of UVB-exposed cells described above was quantified in 150 cells per time point. Data are expressed as average fluorescence/cell (mean±s.d.). (C) Suppression of only the second peak of PARP activation by antioxidants. Cells were incubated for 1 hour with 5000 Units/ml catalase or 20 mM N-acetylcysteine prior to exposure to 1.6 kJ/m² UVB. Cells were then harvested at 15 seconds, 5 minutes or 2 hours. As a control, cells were treated with 100 µM H₂O₂ for 10 minutes. PARP activation was monitored by immunofluorescent detection of pADPr as described for Fig. 1. Data are representative of three experiments with identical results.

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