基于流式细胞术的人衰老细胞整体蛋白质合成评估。

School of Biological Sciences, Seoul National University, Seoul 08826, South Korea.

STAR Protoc. 2021 Sep 14;2(3):100809. doi: 10.1016/j.xpro.2021.100809. eCollection 2021 Sep 17.

Senescent cells constantly experience stressful conditions and restrain their protein translation to cope with it. Here, we present a detailed protocol to measure the rate of global protein synthesis using L-azidohomoalanine (L-AHA)-based click chemistry in human senescent fibroblasts. We optimized several aspects of the procedure, including senescence induction, a flow cytometry analysis of senescent cells, and the duration of L-AHA incorporation. This protocol uses senescent human fibroblasts but can be applied to other types of cells or circumstances. For complete details on the use and execution of this protocol, please refer to Lee et al. (2021).

衰老细胞经常经历应激条件，并抑制其蛋白质翻译以应对它。在这里，我们提出了一种使用 L-叠氮基高丙氨酸 (L-AHA) 基于点击化学的详细方案来测量人衰老成纤维细胞中整体蛋白质合成的速率。我们优化了该程序的几个方面，包括衰老诱导、衰老细胞的流式细胞术分析以及 L-AHA 掺入的持续时间。该方案使用衰老的人成纤维细胞，但可应用于其他类型的细胞或情况。有关此方案的使用和执行的完整详细信息，请参阅 Lee 等人。(2021)。