Flow cytometric analysis of DNA sensitivity to nuclease S1 in UVC-irradiated human fibroblasts

The sensitivity of human fibroblast DNA towards the activity of nuclease S1 was investigated in situ after irradiation of the cells with UVC light. DNA digestion was assessed using flow cytometry by staining the nuclease-resistant DNA fraction with the specific fluorochrome propidium iodide (PI). The results showed a non-linear dependence of DNA digestion on the UVC irradiation dose between 0 and 20 J m-2. About 70% of the UVC-induced DNA sensitivity to nuclease S1 was lost after 30 min of repair. These results suggest that flow cytometry may be useful for assessing the heterogeneity of cell response to UV damage.