GFP-LC3 高内涵分析用于自噬调节剂的筛选。
GFP-LC3 High-Content Assay for Screening Autophagy Modulators.
机构信息
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
Department of Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS, USA.
出版信息
Methods Mol Biol. 2022;2474:83-89. doi: 10.1007/978-1-0716-2213-1_9.
Abstract
Autophagy plays an important role in maintaining cellular homeostasis. Defects in autophagy have been linked to various human diseases, such as cancer, neurodegenerative diseases, and cardiovascular diseases. Therefore, it is useful to develop an assay that can measure the functions of autophagy and also be used to identify autophagy modulators by screening a large number of compounds. This chapter describes a cell-based high content green fluorescent protein (GFP)-LC3 assay using mouse embryonic fibroblasts (MEF) stably expressing GFP-LC3.
摘要
自噬在维持细胞内稳态方面起着重要作用。自噬缺陷与多种人类疾病有关，如癌症、神经退行性疾病和心血管疾病。因此，开发一种能够测量自噬功能的测定法，并通过筛选大量化合物来识别自噬调节剂是很有用的。本章描述了一种基于细胞的高内涵绿色荧光蛋白（GFP）-LC3 测定法，该方法使用稳定表达 GFP-LC3 的小鼠胚胎成纤维细胞（MEF）。
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