Division of Life Science and State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Hong Kong, China.
Bioscience and Biomedical Engineering Thrust, The Hong Kong University of Science and Technology (Guangzhou), Guangzhou, China.
STAR Protoc. 2022 Mar 8;3(1):101227. doi: 10.1016/j.xpro.2022.101227. eCollection 2022 Mar 18.
We present here a protocol to assay the centrosome separation events at late-G2 phase of the cell cycle by immunofluorescence microscopy. We describe the steps required for imaging and measurement of inter-centrosome distance. Here, we use GAS2L1 as an example, but the protocol can be used to test any protein for a role in centrosome separation and cohesion. The steps below are specific for hTERT RPE-1 cell lines, but other adherent cell lines (e.g., U2OS, MRC-5) are also amenable for this protocol. For complete details on the use and execution of this protocol, please refer to Au et al. (2017) and Au et al. (2020).
我们在此提出了一种通过免疫荧光显微镜检测细胞周期晚期 G2 期中心体分离事件的方案。我们描述了进行成像和测量中心体间距离所需的步骤。在这里，我们以 GAS2L1 为例，但该方案可用于测试任何蛋白质在中心体分离和黏附中的作用。以下步骤是针对 hTERT RPE-1 细胞系的，但其他贴壁细胞系（例如 U2OS、MRC-5）也适用于该方案。有关该方案的使用和执行的完整详细信息，请参阅 Au 等人（2017 年）和 Au 等人（2020 年）。
