title: Morphological analysis of autophagy.
abstract: Autophagy is a bulk intracellular degradation process that is ubiquitous in eukaryotic cells and helps to recycle nutrients from catabolites by degrading proteins, lipids, and glycans, including organelles. Since autophagy has divergent physiological roles in cancer, infection, immunity, and other processes, it is important to accurately analyze autophagic activity. In this chapter, we describe methods that can be used to monitor autophagy in cultured mammalian cells by immunostaining and using fluorescently tagged autophagy-related proteins such as GFP- or mRFP-GFP-tandem-tagged proteins as well as electron microscopic methods, including electron tomography and immuno-electron microscopy.
authors.affiliation: Department of Genetics, Graduate School of Medicine, Osaka, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Tabata
authors.forename: Mitsuko
authors.initials: M
authors.lastname: Hayashi-Nishino
authors.forename: Takeshi
authors.initials: T
authors.lastname: Noda
authors.forename: Akitsugu
authors.initials: A
authors.lastname: Yamamoto
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Yoshimori
mesh_terms: D000818:Animals; D001343:Autophagy; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D055032:Electron Microscope Tomography; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006801:Humans; D021621:Imaging, Three-Dimensional; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D016253:Microscopy, Immunoelectron; D008867:Microtomy; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis; D010588:Phagosomes / Q000648:ultrastructure*; D016613:Plastic Embedding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016707:Tissue Fixation; D014162:Transfection; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D049452:Green Fluorescent Proteins; D051907:Lysosomal Membrane Proteins; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; C489116:LAMP1 protein, human; D052118:Lysosomal-Associated Membrane Protein 1; C477012:MAP1LC3A protein, human