Autophagy Detection in Intestinal Stem Cells.

Autophagy Detection in Intestinal Stem Cells.
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肠干细胞中的自噬检测。

DOI:
10.1007/978-1-0716-0747-3_7
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发表时间:
2020
期刊:
Methods Mol Biol.
影响因子:
--
通讯作者:
Ohteki T. (*: equal contribution)
Ohteki T. (*: equal contribution)
中科院分区:
--
文献类型:
--
作者:
Asano J*;Sato T*;Ohteki T. (*: equal contribution)

文献摘要

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自噬是一种溶酶体降解途径,在生理稳态和疾病中起着重要作用。我们之前的研究表明,肠道干细胞(ISCs)的内在自噬对ISC的稳态很重要。本文详细介绍了通过观察GFP-LC3转基因小鼠的自噬体和定量p62蛋白水平来检测ISCs自噬的方法。我们还描述了检测这些细胞中线粒体自噬的方法,分别通过MitoTracker和CellROX溶液分析线粒体跨膜电位和活性氧(ROS)水平。
Autophagy is a lysosomal degradation pathway with important roles in physiological homeostasis and disease. We previously showed that intrinsic autophagy in intestinal stem cells (ISCs) is important for ISC homeostasis. Here we describe the detailed methods for detecting autophagy in ISCs by observing autophagosomes in GFP-LC3 transgenic mice and quantifying the p62 protein levels. We also describe methods for detecting mitophagy in these cells, by analyzing the mitochondrial transmembrane potential and reactive oxygen species (ROS) level by MitoTracker and CellROX solution, respectively.
DOI: 10.1091/mbc.e03-09-0704
发表时间: 2004-03-01
影响因子: 3.3
作者:
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影响因子: 30.3
作者:
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