Pathology Department and Cell Biology Program, Case Western Reserve University, Cleveland, OH 44106, USA.
STAR Protoc. 2021 Jul 7;2(3):100646. doi: 10.1016/j.xpro.2021.100646. eCollection 2021 Sep 17.
can be arrested in metaphase by depleting Cdc20. We describe (1) how to achieve this arrest and verify it, (2) how to label cell surface glycans covalently to distinguish mother from bud, and (3) how to detect the nucleolus and learn that it remains in the mother domain upon arrest. For complete details on the use and execution of this protocol, please refer to Tartakoff et al. (2021), Rai et al. (2017), and Zapanta Rinonos et al. (2014).
可以通过耗尽 Cdc20 使细胞在中期被捕。我们描述了(1)如何实现这种阻滞并进行验证,(2)如何共价标记细胞表面糖基以区分母细胞和芽细胞,以及(3)如何检测核仁并了解到在阻滞时它仍保留在母细胞区域。有关此方案使用和执行的完整详细信息,请参考 Tartakoff 等人(2021 年)、Rai 等人(2017 年)和 Zapanta Rinonos 等人(2014 年)。