Modified LC3 Dot Quantification Method

Tahira Anwar; Eeva-Liisa Eskelinen

doi:10.1007/978-1-0716-2071-7_4

Modified LC3 Dot Quantification Method

Methods Mol Biol. 2022:2445:53-64. doi: 10.1007/978-1-0716-2071-7_4.

Authors

Tahira Anwar¹, Eeva-Liisa Eskelinen²

Affiliations

¹ Molecular and Integrative Biosciences Research Program, University of Helsinki, Helsinki, Finland.
² Institute of Biomedicine, University of Turku, Turku, Finland. eeva-liisa.eskelinen@utu.fi.

PMID: 34972985
DOI: 10.1007/978-1-0716-2071-7_4

Abstract

Since the discovery of autophagy genes and proteins in the early1990s, numerous previously unknown physiological and pathological functions have been discovered for autophagy. At the same time, precise monitoring of autophagy has become important, and western blotting and fluorescence microscopy of the marker protein LC3 is widely used for this purpose. Here, we describe a modification of the widely used method, number of LC3 dots per cell. This protocol provides the proportion of vesicular LC3 staining over the total LC3 staining in the same cell. The approach is well suitable for quantification of endogenous LC3.

Keywords: Autophagy; Cell profiler; Fluorescence microscopy; Image analysis; Immunofluorescence; LC3.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Autophagy / physiology
Blotting, Western
Microscopy, Fluorescence
Microtubule-Associated Proteins* / metabolism
Tomography, Optical*

Substances

Microtubule-Associated Proteins