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. 2009:452:1-12.
doi: 10.1016/S0076-6879(08)03601-X.

Monitoring autophagy in mammalian cultured cells through the dynamics of LC3

Shunsuke Kimura  1 , Naonobu Fujita, Takeshi Noda, Tamotsu Yoshimori
Affiliations

Monitoring autophagy in mammalian cultured cells through the dynamics of LC3

Shunsuke Kimura et al. Methods Enzymol. 2009.

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.

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