A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts

Summary: Autophagy measurement has been challenging due to the transient nature of autophagy vesicles, in which degradation of cargo occurs. Here, we present a protocol to monitor starvation-induced autophagy using a live high-throughput microscopy system in a fast and automated manner without the n...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Ada Nowosad, Arnaud Besson
Formato: article
Lenguaje:EN
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://doaj.org/article/4fd57986613f436d96f10c6836e8d563
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
id oai:doaj.org-article:4fd57986613f436d96f10c6836e8d563
record_format dspace
spelling oai:doaj.org-article:4fd57986613f436d96f10c6836e8d5632021-11-18T04:51:44ZA high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts2666-166710.1016/j.xpro.2021.100966https://doaj.org/article/4fd57986613f436d96f10c6836e8d5632021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721006729https://doaj.org/toc/2666-1667Summary: Autophagy measurement has been challenging due to the transient nature of autophagy vesicles, in which degradation of cargo occurs. Here, we present a protocol to monitor starvation-induced autophagy using a live high-throughput microscopy system in a fast and automated manner without the need for sample preparation. We provide a detailed protocol describing the generation of turboGFP-LC3B expressing mouse embryonic fibroblasts (MEFs), the measurement of autophagy over time and the analysis of data.For complete details on the use and execution of this protocol, please refer to Nowosad et al. (2020, 2021).Ada NowosadArnaud BessonElsevierarticleCell BiologyCell-based AssaysHigh Throughput ScreeningMicroscopyScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100966- (2021)
institution DOAJ
collection DOAJ
language EN
topic Cell Biology
Cell-based Assays
High Throughput Screening
Microscopy
Science (General)
Q1-390
spellingShingle Cell Biology
Cell-based Assays
High Throughput Screening
Microscopy
Science (General)
Q1-390
Ada Nowosad
Arnaud Besson
A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts
description Summary: Autophagy measurement has been challenging due to the transient nature of autophagy vesicles, in which degradation of cargo occurs. Here, we present a protocol to monitor starvation-induced autophagy using a live high-throughput microscopy system in a fast and automated manner without the need for sample preparation. We provide a detailed protocol describing the generation of turboGFP-LC3B expressing mouse embryonic fibroblasts (MEFs), the measurement of autophagy over time and the analysis of data.For complete details on the use and execution of this protocol, please refer to Nowosad et al. (2020, 2021).
format article
author Ada Nowosad
Arnaud Besson
author_facet Ada Nowosad
Arnaud Besson
author_sort Ada Nowosad
title A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts
title_short A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts
title_full A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts
title_fullStr A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts
title_full_unstemmed A high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts
title_sort high-throughput protocol for monitoring starvation-induced autophagy in real time in mouse embryonic fibroblasts
publisher Elsevier
publishDate 2021
url https://doaj.org/article/4fd57986613f436d96f10c6836e8d563
work_keys_str_mv AT adanowosad ahighthroughputprotocolformonitoringstarvationinducedautophagyinrealtimeinmouseembryonicfibroblasts
AT arnaudbesson ahighthroughputprotocolformonitoringstarvationinducedautophagyinrealtimeinmouseembryonicfibroblasts
AT adanowosad highthroughputprotocolformonitoringstarvationinducedautophagyinrealtimeinmouseembryonicfibroblasts
AT arnaudbesson highthroughputprotocolformonitoringstarvationinducedautophagyinrealtimeinmouseembryonicfibroblasts
_version_ 1718424971603083264