Universität zu Köln

Springhorn, Alexander and Hoppe, Thorsten ORCID: 0000-0002-4734-9352 (2019). Western blot analysis of the autophagosomal membrane protein LGG-1/LC3 in Caenorhabditis elegans. In: Methods in Enzymology, S. 319 - 337. SAN DIEGO: ELSEVIER ACADEMIC PRESS INC. ISBN 978-0-12-818667-1

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Abstract

Autophagy is being studied intensively in Caenorhabditis elegans in the context of protein homeostasis and aging. However, in contrast to the yeast and mammalian autophagosomal membrane proteins Atg8 and LC3, lipidation of the C. elegans ortholog LGG-1 with phosphatidylethanolamine has rarely been investigated by western blotting. We attribute this shortcoming to technical problems with separating the non-lipidated from the lipidated LGG-1 protein by gel electrophoresis. Our new protocol for Western blot analysis is applicable for both the detection of transgenic and endogenous LGG-1 proteins and provides a quantifiable method to assess autophagic flux. As a proof of principle, we use this assay to analyze the role of the transcriptional master regulator HLH-30/TFEB in starvation-induced autophagy.

Item Type:	Book Section, Proceedings Item or annotation in a legal commentary
Creators:	Creators Email ORCID ORCID Put Code Springhorn, Alexander UNSPECIFIED UNSPECIFIED UNSPECIFIED Hoppe, Thorsten UNSPECIFIED orcid.org/0000-0002-4734-9352 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-160239
DOI:	10.1016/bs.mie.2018.12.034
Title of Book:	Methods in Enzymology
Series Name:	Methods Enzymol.
Volume:	619
Page Range:	S. 319 - 337
Date:	2019
Publisher:	ELSEVIER ACADEMIC PRESS INC
Place of Publication:	SAN DIEGO
ISSN:	0076-6879
ISBN:	978-0-12-818667-1
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Institute for Genetics
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language LC3; GENES; ATG9; LONGEVITY; TRANSPORT; SURVIVAL Multiple languages Biochemical Research Methods; Biochemistry & Molecular Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/16023