Abstract
We describe the use of flow cytometric methods to analyze DNA damage. Cultured human cells treated with sulfur mustard (HD) were analyzed for DNA strand breakage by three methods. First, we used cell cycle analysis by propidium iodide (PI) uptake for determination of cell cycle disruption and DNA fragmentation after HD exposure. Second, a terminal deoxynucleotidyl transferase (TdT) assay allowed for quantitation of DNA strand breaks resulting from treatment with HD. Last, we used immunostaining of the guanine N7 adducts (the primary DNA alkylation product resulting from HD exposure) as a measure of DNA repair. With these methods, it was possible to describe both concentration- and time-dependent effects of HD-exposure in human epithelial cells. We describe applications of these methods to future studies aimed at reducing DNA damage resulting from HD exposure.
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