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Fig. 4. Colocalization of hybridization signals of homologous chromosome bands in human fibroblasts. The percentage of cells with one hybridization signal for the homologues of either chromosome 8 or 9 was analyzed manually. (A) The analysis was performed on confluent cells treated with 4 µM MMC for 1 hour and fixed either immediately (recovery time 0 hours) or after a recovery time of 20 hours. Solvent controls were included for both time points. Each bar represents the average percentage obtained from at least three independent experiments. In each experiment 500 cells were analyzed. The error bars are standard deviations (s.d.) of the mean values. A significantly increased colocalization was evident for chromosome 9 in the treated cells (P<0.05, Student t-test) and no difference was observed between 0 hour and 20 hour recovery. (B) The analysis was performed on cycling cells treated with 4 µM MMC for 1 hour, 24 hours after they were released from confluency by subcultivation to reduced cell density. The cells were fixed directly after the treatment. Since BrdU was present during treatment, BrdU-positive cells must have been treated in the S-phase of their cell cycle. 500 BrdU-positive and -negative cells were analyzed. A significant increase of the number of cells containing one hybridization signal for chromosome 9 was observed in MMC-treated cells (P<0.05, Wilson's method for difference in proportions).
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