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Fig. 6. Mimosine-induced DNA damage is cell cycle dependent. (A) Quiescent EJ30 cells were incubated in 500 µM mimosine for 24 hours, fixed, treated with RNase A, and stained for DNA (red) and for {gamma}-H2AX (green). (B) Proliferating EJ30 cells were incubated for 24 hours and stained as in A. (C) EJ30 cells were arrested in mitosis by a 24-hour nocodazole treatment, released by replacing the medium, and then subjected to a treatment of 500 µM mimosine or 5 µg/ml aphidicolin in intervals of 4-8, 8-12 and 12-16 hours post release. The percentage of cells showing focal {gamma}-H2AX staining were determined by immunofluorescence microscopy. (D) Flow-cytometry analysis of the DNA content of untreated control cells at the indicated time points after release from nocodazole as used in C. The percentages of cells that incorporated BrdU in the 15 minutes before preparation are shown in each panel.





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