Autophagy detection in intestinal stem cells

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.