Chapter 1 Monitoring Autophagy in Mammalian Cultured Cells through the Dynamics of LC3

Shunsuke Kimura, Naonobu Fujita, Takeshi Noda, Tamotsu Yoshimori

Research output: Contribution to journalReview article

175 Citations (Scopus)

Abstract

In this chapter, we introduce several methods that rely on the analysis of LC3, a versatile marker protein of autophagic structures in mammalian cultured cells. The appearance of LC3-positive puncta is indicative of the induction of autophagy, and it is observed either by immunofluorescence or by GFP-based microscopy. The maturation process by which autophagosomes are converted into autolysosomes can be monitored by the GFP and RFP tandemly tagged LC3 (tfLC3) method. Lysosomal turnover of LC3 is a good index of the proceeding of autophagy and can be assessed by Western blotting. These methods will provide a relatively easy assessment of autophagy, and the details of the procedure will be described along with possible pitfalls.

Original languageEnglish
Pages (from-to)1-12
Number of pages12
JournalMethods in Enzymology
Volume451
Issue numberC
DOIs
Publication statusPublished - 2009 Feb 9
Externally publishedYes

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Autophagy
Cultured Cells
Microscopic examination
Cells
Monitoring
Proteins
Fluorescent Antibody Technique
Microscopy
Western Blotting

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Chapter 1 Monitoring Autophagy in Mammalian Cultured Cells through the Dynamics of LC3. / Kimura, Shunsuke; Fujita, Naonobu; Noda, Takeshi; Yoshimori, Tamotsu.

In: Methods in Enzymology, Vol. 451, No. C, 09.02.2009, p. 1-12.

Research output: Contribution to journalReview article

Kimura, Shunsuke ; Fujita, Naonobu ; Noda, Takeshi ; Yoshimori, Tamotsu. / Chapter 1 Monitoring Autophagy in Mammalian Cultured Cells through the Dynamics of LC3. In: Methods in Enzymology. 2009 ; Vol. 451, No. C. pp. 1-12.
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