Monitoring Autophagy with GFP-LC3 Reporter

Lidong Sun; Suli Lv; Tanjing Song

doi:10.1007/7651_2023_501

Monitoring Autophagy with GFP-LC3 Reporter

Methods Mol Biol. 2023 Oct 28. doi: 10.1007/7651_2023_501. Online ahead of print.

Authors

Lidong Sun^{1

2}, Suli Lv³, Tanjing Song^{4

5}

Affiliations

¹ Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. LidongSun@hust.edu.cn.
² Cell Architecture Research Institute, Huazhong University of Science and Technology, Wuhan, Hubei, China. LidongSun@hust.edu.cn.
³ Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
⁴ Department of Biochemistry and Molecular Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. SongT@hust.edu.cn.
⁵ Cell Architecture Research Institute, Huazhong University of Science and Technology, Wuhan, Hubei, China. SongT@hust.edu.cn.

PMID: 37889422
DOI: 10.1007/7651_2023_501

Abstract

Autophagy is a key process that maintains cellular homeostasis. Autophagy contributes to various physiological and pathophysiological processes. Development of methodologies for autophagy detection has greatly facilitated the research on autophagy. Among these methodologies, GFP-LC3 reporter has been popularly used in the literature. In this chapter, we will detail step-by-step the GFP-LC3 reporter protocol we have adapted in our lab. This protocol begins with the generation of lentivirus expressing GFP-LC3. Then, the cells are transduced with titrated virus. After selecting the positive cells, single colonies are isolated, characterized, validated, and used in further study.

Keywords: Autophagosome; Autophagy; GFP; LC3; Lysosome.