Chapter 9 Flow Cytometric Analysis of Autophagy in Living Mammalian Cells

Methods in Enzymology

Volumn 451, Issue C, 2009, Pages 131-141

  Shvets, Elena ^a   Elazar, Zvulun ^a  

^a WEIZMANN INSTITUTE OF SCIENCE   (Israel)

Author keywords

[No Author keywords available]

Indexed keywords

GREEN FLUORESCENT PROTEIN; MICROTUBULE ASSOCIATED PROTEIN; AMINO ACID; LEUCINE; PHOSPHATIDYLETHANOLAMINE; RAPAMYCIN; TUNICAMYCIN; UBIQUITIN;

ANIMAL; ARTICLE; AUTOPHAGY; CHO CELL; CRICETULUS; FLOW CYTOMETRY; GENETICS; HAMSTER; METABOLISM; METHODOLOGY; PHYSIOLOGY; ACCURACY; BIOCHEMISTRY; CELL POPULATION; CELL SIZE; FLUORESCENCE ACTIVATED CELL SORTING; HUMAN; IMMUNOFLUORESCENCE MICROSCOPY; MAMMAL CELL; NONHUMAN; PHAGOSOME; PRIORITY JOURNAL; PROTEIN EXPRESSION; QUALITATIVE ANALYSIS; QUANTITATIVE ANALYSIS; REVIEW; SIGNAL TRANSDUCTION;

ANIMALS; AUTOPHAGY; CHO CELLS; CRICETINAE; CRICETULUS; FLOW CYTOMETRY; GREEN FLUORESCENT PROTEINS; MICROTUBULE-ASSOCIATED PROTEINS;

MAMMALIA;

EID: 59249105119     PISSN: 00766879     EISSN: None     Source Type: Book Series    
DOI: 10.1016/S0076-6879(08)03609-4     Document Type: Review

References (19)

1. Bampton E.T., Goemans C.G., Niranjan D., Mizushima N., and Tolkovsky A.M. The dynamics of autophagy visualized in live cells: From autophagosome formation to fusion with endo/lysosomes. Autophagy 1 (2005) 23-36
2. Blommaart E.F., Krause U., Schellens J.P., Vreeling-Sindelarova H., and Meijer A.J. The phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 inhibit autophagy in isolated rat hepatocytes. Eur. J. Biochem. 243 (1997) 240-246
3. Boland B., and Nixon R.A. Neuronal macroautophagy: From development to degeneration. Mol. Aspects. Med. 27 (2006) 503-519
4. Campbell R.E., Tour O., Palmer A.E., Steinbach P.A., Baird G.S., Zacharias D.A., and Tsien R.Y. A monomeric red fluorescent protein. Proc. Natl. Acad. Sci. USA 99 (2002) 7877-7882
5. Fass E., Shvets E., Degani I., Hirschberg K., and Elazar Z. Microtubules support production of starvation-induced autophagosomes but not their targeting and fusion with lysosomes. J. Biol. Chem. 281 (2006) 36303-36316
6. Huang J., and Klionsky D.J. Autophagy and human disease. Cell Cycle 6 (2007) 1837-1849
7. Kabeya Y., Mizushima N., Ueno T., Yamamoto A., Kirisako T., Noda T., Kominami E., Ohsumi Y., and Yoshimori T. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 19 (2000) 5720-5728
8. Kabeya Y., Mizushima N., Yamamoto A., Oshitani-Okamoto S., Ohsumi Y., and Yoshimori T. LC3, GABARAP and GATE16 localize to autophagosomal membra on form-II formation. J. Cell Sci. 117 (2004) 2805-2812
9. Kimura S., Noda T., and Yoshimori T. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 3 (2007) 452-460
10. Klionsky D.J. The molecular machinery of autophagy: Unanswered questions. J. Cell Sci. 118 (2005) 7-18
11. Kneen M., Farinas J., Li Y., and Verkman A.S. Green fluorescent protein as a noninvasive intracellular pH indicator. Biophys. J. 74 (1998) 1591-1599
12. Kuma A., Matsui M., and Mizushima N. LC3, an autophagosome marker, can be incorporated into protein aggregates independent of autophagy: Caution in the interpretation of LC3 localization. Autophagy 3 (2007) 323-328
13. Mizushima A., and Kuma. Autophagosomes in GFP-LC3 Transgenic Mice. Methods Mol. Biol. 445 (2008) 119-124
14. Mizushima N., Yamamoto A., Matsui M., Yoshimori T., and Ohsumi Y. In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol. Biol. Cell 15 (2004) 1101-1111
15. Mortimore G.E., Poso A.R., Kadowaki M., and Wert Jr. J.J. Multiphasic control of hepatic protein degradation by regulatory amino acids. General features and hormonal modulation. J. Biol. Chem. 262 (1987) 16322-16327
16. Ohsumi Y. Molecular dissection of autophagy: two ubiquitin-like systems. Nat. Rev. Mol. Cell Biol. 2 (2001) 211-216
17. Shintani, and Klionsky. Autophagy in health and disease: A double-edged sword. Science 306 (2004) 990-995
18. Shvets E., Fass E., and Elazar Z. Utilizing flow cytometry to monitor autophagy in living mammalian cells. Autophagy 4 (2008) 621-628
19. Tanida I., Minematsu-Ikeguchi N., Ueno T., and Kominami E. Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 1 (2005) 84-91