HeLa/RFP-H2B cell lines bearing the K209A or K209M mutant were synchronized by T/N block and release as described above. Harvested cells were treated with 0.075 M KCl hypotonic buffer at a 37°C water bath for 15 min, followed by fixation with methanol:acetic acid (3:1, v/v). Cell suspensions were dropped onto cleaned slides and air-dried. The slides were stained with a diluted Giemsa solution for 10 min and then were washed by water and air-dried. Images were acquired by Leica DM1000 and analyzed using Leica Application Suite V3 software.

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