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 articlepeer-review

    189 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

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

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