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Autophagy Detection in Intestinal Stem Cells

Jumpei Asano^1,2, Taku Sato¹, Toshiaki Ohteki³

¹Department of Biodefense Research, Medical Research Institute, Tokyo Medical and Dental University (TMDU), Tokyo, Japan.

Methods in Molecular Biology (Clifton, N.J.)

|July 25, 2020

Abstract:

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.

Keywords:

Atg5 Autophagosome Autophagy GFP-LC3 Intestinal epithelial cells (IECs)Intestinal stem cells (ISCs)Lgr5 Mitochondria Reactive oxygen species (ROS)p62

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Keywords:

Atg5 Autophagosome Autophagy GFP-LC3 Intestinal epithelial cells (IECs)Intestinal stem cells (ISCs)Lgr5 Mitochondria Reactive oxygen species (ROS)p62

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