Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence

Senescent cells accumulation is associated with aging and age-related diseases, and recent findings suggest that autophagy, the activity of the intracellular degradation system, decreases during senescence. In this protocol, we detail steps to induce cellular senescence in response to DNA damage, ev...

Descripción completa

Detalles Bibliográficos
Autores principales: Yamamoto-Imoto, Hitomi, Hara, Eiji, Nakamura, Shuhei, Yoshimori, Tamotsu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9283947/
https://www.ncbi.nlm.nih.gov/pubmed/35819884
http://dx.doi.org/10.1016/j.xpro.2022.101539
_version_ 1784747446527590400
author Yamamoto-Imoto, Hitomi
Hara, Eiji
Nakamura, Shuhei
Yoshimori, Tamotsu
author_facet Yamamoto-Imoto, Hitomi
Hara, Eiji
Nakamura, Shuhei
Yoshimori, Tamotsu
author_sort Yamamoto-Imoto, Hitomi
collection PubMed
description Senescent cells accumulation is associated with aging and age-related diseases, and recent findings suggest that autophagy, the activity of the intracellular degradation system, decreases during senescence. In this protocol, we detail steps to induce cellular senescence in response to DNA damage, evaluate the senescent state using SA-β-gal staining and western blot for p21, LAMP1, and Lamin B1, and detect autophagy via LC3 western blotting. This protocol can be used in most cell lines and for various types of senescent cells. For complete details on the use and execution of this protocol, please refer to Yamamoto-Imoto et al. (2022).
format Online
Article
Text
id pubmed-9283947
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-92839472022-07-16 Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence Yamamoto-Imoto, Hitomi Hara, Eiji Nakamura, Shuhei Yoshimori, Tamotsu STAR Protoc Protocol Senescent cells accumulation is associated with aging and age-related diseases, and recent findings suggest that autophagy, the activity of the intracellular degradation system, decreases during senescence. In this protocol, we detail steps to induce cellular senescence in response to DNA damage, evaluate the senescent state using SA-β-gal staining and western blot for p21, LAMP1, and Lamin B1, and detect autophagy via LC3 western blotting. This protocol can be used in most cell lines and for various types of senescent cells. For complete details on the use and execution of this protocol, please refer to Yamamoto-Imoto et al. (2022). Elsevier 2022-07-11 /pmc/articles/PMC9283947/ /pubmed/35819884 http://dx.doi.org/10.1016/j.xpro.2022.101539 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Yamamoto-Imoto, Hitomi
Hara, Eiji
Nakamura, Shuhei
Yoshimori, Tamotsu
Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence
title Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence
title_full Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence
title_fullStr Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence
title_full_unstemmed Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence
title_short Measurement of autophagy via LC3 western blotting following DNA-damage-induced senescence
title_sort measurement of autophagy via lc3 western blotting following dna-damage-induced senescence
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9283947/
https://www.ncbi.nlm.nih.gov/pubmed/35819884
http://dx.doi.org/10.1016/j.xpro.2022.101539
work_keys_str_mv AT yamamotoimotohitomi measurementofautophagyvialc3westernblottingfollowingdnadamageinducedsenescence
AT haraeiji measurementofautophagyvialc3westernblottingfollowingdnadamageinducedsenescence
AT nakamurashuhei measurementofautophagyvialc3westernblottingfollowingdnadamageinducedsenescence
AT yoshimoritamotsu measurementofautophagyvialc3westernblottingfollowingdnadamageinducedsenescence

Ejemplares similares