Comet assay was performed as previously described (49). Briefly, cells were treated as indicated and then mixed gently with premelted low-temperature–melting agarose at a volume ratio of 1:1 (v/v) and spread on glass slides. The slides were then submerged in precooled lysis buffer at 4°C for 90 min. After rinsing, the slides were electrophoresed in running buffer [1 mM Na2EDTA and 300 mM NaOH (pH 13.0)] at 1.0 V/cm for 20 min and then stained with propidium iodide (5 μg/ml). Fluorescent images of more than 100 nuclei were captured under an Olympus FV1000-IX81 confocal microscope (Tokyo, Japan).

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