Figure 8.

Phosphorylation at Ser1213 and S1525 controls the timing of TOP2A recruitment at centromeres. U2OS cells were tranfected with constructs expressing either WT, S1213A/S1525A or S1213D/S1525D FLAG-tagged TOP2A. After 48 h, cell extracts were prepared and analysed by western blotting (A) or cells were treated with 10 μM EdU and 50 μg/ml ICRF-187 for 15 min prior to pre-extraction, fixation and immunofluorecence microscopy analysis (B). Representative images of early S-phase cells are shown. (C) Quantification was performed as in Figures 6 and 7. ***P < 0.001, Student's t-test. Scale bar = 10 μm.