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...

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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
Descripción
Sumario: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).

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