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Fig. 3. Treatment with iron chelators causes DNA breaks. (A) EJ30 cells were treated with the indicated compounds for 24 hours, fixed, treated with RNase A and stained with propidium iodide for DNA (red) and with antibodies for {gamma}-H2AX (green). Merged images are shown. (B) HeLa cells were subjected to treatments exactly as in A and the generated DNA breaks were analysed by alkaline-agarosegel electrophoresis. Lanes 1 and 7, untreated control; 2, thymidine; 3, aphidicolin; 4, mimosine; 5, ciclopirox olamine; 6, bipyridyl; 8, etoposide; 9, 1 µg/ml Bleocin; 10, 10 µg/ml Bleocin. The marker (M) is a 1 kb ladder (Roche). (C) Samples 1-4 from B were subjected to neutral pulsed-field-gel electrophoresis to assay for the presence of DSBs. The markers cover the 0.1-200 kb (M1) or 50-1000 kb (M2) size-range.