Centre de Recherche des Cordeliers, Université de Paris, Sorbonne Université, Inserm U1138, Institut Universitaire de France, Paris, France.
Cell biology and metabolomics platforms, Gustave Roussy Cancer Center, Villejuif, France.
Methods Mol Biol. 2021;2267:217-226. doi: 10.1007/978-1-0716-1217-0_15.
Mitotic catastrophe is an oncosuppressive mechanism that drives cells toward senescence or death when an error occurs during mitosis. Eukaryotic cells have developed adaptive signaling pathways to cope with stress. The phosphorylation on serine 51 of the eukaryotic translation initiation factor (eIF2α) is a highly conserved event in stress responses, including the one that is activated upon treatment with mitotic catastrophe inducing agents, such as microtubular poisons or actin blockers. The protocol described herein details a method to quantify the phosphorylation of eIF2α by high-throughput immunofluorescence microscopy. This method is useful to capture the 'integrated stress response', which is characterized by eIF2α phosphorylation in the context of mitotic catastrophe.
有丝分裂灾难是一种抑瘤机制，当有丝分裂过程中出现错误时，它会促使细胞衰老或死亡。真核细胞已经开发出适应性信号通路来应对压力。真核翻译起始因子 (eIF2α) 丝氨酸 51 的磷酸化是应激反应中的一个高度保守事件，包括用有丝分裂灾难诱导剂（如微管毒素或肌动蛋白阻断剂）处理时激活的反应。本文所述的方案详细介绍了一种通过高通量免疫荧光显微镜定量测定 eIF2α 磷酸化的方法。该方法可用于捕获“综合应激反应”，其特征是有丝分裂灾难背景下的 eIF2α 磷酸化。
