Institute of Biochemistry 2, Faculty of Medicine, Goethe University Frankfurt, Frankfurt am Main, Germany.
Buchmann Institute for Molecular Life Sciences, Goethe University Frankfurt, Frankfurt am Main, Germany.
Methods Mol Biol. 2023;2706:215-224. doi: 10.1007/978-1-0716-3397-7_16.
Autophagy is a cellular process implicated in the renewal of cellular components and the maintenance of cellular hemostasis and therefore associated with various types of diseases. In addition, autophagy belongs to the stress response pathways and is frequently activated by chemical compounds harboring characteristics of cell toxicity. High-throughput screens analyzing autophagy flux are therefore applied in both, the field of compound identification for targeting autophagy and compound characterization for analyzing compound toxicity. In this chapter, we describe a live-cell, fluorescent-based, high-throughput screening method in 384-well format for the fast and accurate measurement of autophagy flux over time suitable for academic research, pharmacological applications, and drug discovery.
自噬是一种与多种疾病相关的细胞过程，涉及细胞成分的更新和细胞稳态的维持。此外，自噬属于应激反应途径，经常被具有细胞毒性特征的化合物激活。因此，分析自噬通量的高通量筛选既应用于靶向自噬的化合物鉴定领域，也应用于分析化合物毒性的化合物特性分析领域。在本章中，我们描述了一种基于荧光的、适用于 384 孔板的活细胞高通量筛选方法，可快速、准确地测量随时间推移的自噬通量，适用于学术研究、药理学应用和药物发现。
