Gillespie David A F, Walker Mark
CR-UK Beatson Laboratories, Beatson Institute for Cancer Research, Garscube Estate, Bearsden, Glasgow G61 1BD.
Subcell Biochem. 2006;40:355-8. doi: 10.1007/978-1-4020-4896-8_25.
Chromosome condensation during mitosis is associated with phosphorylation of histone H3 at serine 10 (pS10H3). Detection of pS10H3 phosphorylation using phospho-specific antibodies in combination with DNA content flow cytometry provides a rapid and accurate means of determining the percentage of mitotic cells in a population. Using the spindle poison nocodazole to trap mitotic cells it is possible to determine the rate at which cells accumulate in mitosis with time. By comparing the rates of mitotic accumulation before and after DNA damage it is possible to gauge the efficiency with which the G2 checkpoint blocks entry to mitosis. Because DNA content is measured simultaneously with pS10H3 fluorescence this method can also be used to identify cells which enter mitosis with incompletely replicated DNA as a result of S-M checkpoint failure during DNA synthesis inhibition.
有丝分裂期间的染色体凝聚与组蛋白H3丝氨酸10位点的磷酸化（pS10H3）相关。使用磷酸特异性抗体结合DNA含量流式细胞术检测pS10H3磷酸化，为确定群体中有丝分裂细胞的百分比提供了一种快速准确的方法。使用纺锤体毒素诺考达唑捕获有丝分裂细胞，可以确定细胞随时间在有丝分裂中积累的速率。通过比较DNA损伤前后有丝分裂积累的速率，可以评估G2期检查点阻止进入有丝分裂的效率。由于DNA含量与pS10H3荧光同时测量，该方法还可用于识别由于DNA合成抑制期间S-M检查点功能障碍而携带未完全复制DNA进入有丝分裂的细胞。
