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Protocol

Autophagosome immunoisolation from GFP-LC3B mouse tissue

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Pages 341-346 | Received 31 Jan 2018, Accepted 17 Oct 2018, Published online: 30 Oct 2018

References

  • Yang Z, Klionsky DJ. Mammalian autophagy: core molecular machinery and signaling regulation. Curr Opin Cell Biol. 2010;22:124–131.
  • Yoshimori T. Autophagy: a regulated bulk degradation process inside cells. Biochem Biophys Res Commun. 2004;313:453–458.
  • Deter RL. Electron microscopic evaluation of subcellular fractions obtained by ultracentrifugation. In: Hayat MA, editor. Principles and techniques of electron microscopy. New York: Van Nostrand Reinhold Co; 1973. p. 199–235.
  • Marzella L, Ahlberg J, Glaumann H. Isolation of autophagic vacuoles from rat liver: morphological and biochemical characterization. J Cell Biol. 1982;93:144–154.
  • Strømhaug PE, Berg TO, Fengsrud M, et al. Purification and characterization of autophagosomes from rat hepatocytes. Biochem J. 1998;335:217–224.
  • Seglen PO, Brinchmann MF. Purification of autophagosomes from rat hepatocytes. Autophagy. 2010;6:542–547.
  • Chen X, Li LJ, Zheng XY, et al. Isolation of autophagosome subpopulations after induction of autophagy by calcium. Biochem Cell Biol. 2015;93:180–184.
  • Gao W, Kang JH, Liao Y, et al. Biochemical isolation and characterization of the tubulovesicular LC3-positive autophagosomal compartment. J Biol Chem. 2010;285:1371–1383.
  • Yao J, Jia L, Feathers K, et al. Autophagy-mediated catabolism of visual transduction proteins prevents retinal degeneration. Autophagy. 2016;12:2439–2450.
  • Kabeya Y, Mizushima N, Ueno T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Embo J. 2000;19:5720–5728.
  • Mizushima N. Methods for monitoring autophagy. Int J Biochem Cell Biol. 2004;36:2491–2502.
  • Mizushima N, Kuma A. Autophagosomes in GFP-LC3 transgenic mice. Meth Mol Biol. 2008;445:119–124.
  • Mizushima N, Yamamoto A, Matsui M, et al. In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol Biol Cell. 2004;15:1101–1111.
  • Matsumoto G, Wada K, Okuno M, et al. Serine 403 phosphorylation of p62/SQSTM1 regulates selective autophagic clearance of ubiquitinated proteins. Mol Cell. 2011;44:279–289.
  • Haspel J, Shaik RS, Ifedigbo E, et al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011;7:629–642.
  • Yao J, Qiu Y, Frontera E, et al. Inhibiting autophagy reduces retinal degeneration caused by protein misfolding. Autophagy. 2018;14:1226–1238.
  • Storrie B, Madden EA. Isolation of subcellular organelles. Methods Enzymol. 1990;182:203–225.
  • Hara T, Nakamura K, Matsui M, et al. Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice. Nature. 2006;441:885–889.
  • Komatsu M, Waguri S, Ueno T, et al. Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice. J Cell Biol. 2005;169:425–434.
  • Komatsu M, Waguri S, Chiba T, et al. Loss of autophagy in the central nervous system causes neurodegeneration in mice. Nature. 2006;441:880–884.
  • Mizushima N, Kuma S. Autophagosomes in GFP-LC3 transgenic mice. Methods Mol Biol. 2008;445:119–124.
  • Kuma A, Matsui M, Mizushima N. LC3, an autophagosome marker, can be incorporated into protein aggregates independent of autophagy; caution in the interpretation of LC3 localization. Autophagy. 2007;3:323–328.
  • Esteban-Martinez L, Boya P. Autophagic flux determination in vivo and ex vivo. Methods. 2015;75:79–86.

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