title: Quantifying Autophagic Structures in Mammalian Cells Using Confocal Microscopy.
abstract: Autophagy relies on the sequential, hierarchical association of proteins with phagophores, and forming autophagosomes to allow completion of the process. Additionally, the trafficking of the unique transmembrane autophagy-related protein ATG9 is vital for autophagy progression. In this chapter, we discuss methods to monitor autophagosome number using confocal microscopy, by following the association of different autophagosomal markers with the phagophore and completed autophagosome. We also discuss methods to monitor the trafficking of ATG9 in mammalian cells under starvation conditions.
authors.affiliation: Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom. Electronic address: christopheralamb101@gmail.com.
authors.forename: C A
authors.initials: CA
authors.lastname: Lamb
authors.affiliation: Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom.
authors.forename: J
authors.initials: J
authors.lastname: Joachim
authors.affiliation: Molecular Cell Biology of Autophagy Group, The Francis Crick Institute, London, United Kingdom. Electronic address: sharon.tooze@crick.ac.uk.
authors.forename: S A
authors.initials: SA
authors.lastname: Tooze
mesh_terms: D000818:Animals; D000071182:Autophagosomes / Q000648:ultrastructure*; D000071183:Autophagy-Related Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D008322:Mammals; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism
chemical_list: C574794:ATG9A protein, human; D000071183:Autophagy-Related Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D033921:Vesicular Transport Proteins
keywords: ATG9; Autophagosome; Autophagy; Imaris; LC3; Live cell imaging