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Figure 6


Fig. 6. ssDNA focus formation is stimulated by S-phase progression. (A) GM-847 cells were transfected with the indicated siRNAs, and BrdU was immediately added to the growth medium. At 24 hours following transfection, BrdU was removed from the cells and mimosine or HU were added. 24 hours thereafter, drugs were removed and the cells were incubated in normal growth medium until fixation at the indicated time-points. (B) ssDNA focus formation following release from mimosine block. The upper panel shows laser scanning flow cytometry analysis of TO-PRO-3-labeled cells. The lower panel shows quantitative determination of cells containing BrdU-labeled ssDNA foci. Bars represent the percentage of cells containing ssDNA foci. For each sample, more than 200 cells were evaluated. Data represent the average of two independent experiments ± s.d. (C) ssDNA focus formation following release from HU block. The same experiment as in B, except HU was used instead of mimosine. `C' indicates samples where cells have not been treated with mimosine or HU.





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