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Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice

Innate immunity is the first layer of defense against infection in mammals and is tightly regulated. We monitored cGAS/STING signaling upon ISD90 or 2′,3′-cGAMP stimulation in EA.hy926 cells by western blotting, RT-PCR, and ELISA analyses to reveal signaling activation and IFNβ production. In additi...

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Detalles Bibliográficos
Autores principales: Wang, Ying, Ma, Zhe, Damania, Blossom, Liu, Pengda
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757363/
https://www.ncbi.nlm.nih.gov/pubmed/33377065
http://dx.doi.org/10.1016/j.xpro.2020.100171
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author Wang, Ying
Ma, Zhe
Damania, Blossom
Liu, Pengda
author_facet Wang, Ying
Ma, Zhe
Damania, Blossom
Liu, Pengda
author_sort Wang, Ying
collection PubMed
description Innate immunity is the first layer of defense against infection in mammals and is tightly regulated. We monitored cGAS/STING signaling upon ISD90 or 2′,3′-cGAMP stimulation in EA.hy926 cells by western blotting, RT-PCR, and ELISA analyses to reveal signaling activation and IFNβ production. In addition, we also include an HSV-1 infected mouse model to further reveal procedures in analyzing cGAS/STING signaling in mice. This protocol could be applied to studies focusing on cell culture or mouse models to investigate cGAS/STING signaling. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2020).
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spelling pubmed-77573632020-12-28 Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice Wang, Ying Ma, Zhe Damania, Blossom Liu, Pengda STAR Protoc Protocol Innate immunity is the first layer of defense against infection in mammals and is tightly regulated. We monitored cGAS/STING signaling upon ISD90 or 2′,3′-cGAMP stimulation in EA.hy926 cells by western blotting, RT-PCR, and ELISA analyses to reveal signaling activation and IFNβ production. In addition, we also include an HSV-1 infected mouse model to further reveal procedures in analyzing cGAS/STING signaling in mice. This protocol could be applied to studies focusing on cell culture or mouse models to investigate cGAS/STING signaling. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2020). Elsevier 2020-11-14 /pmc/articles/PMC7757363/ /pubmed/33377065 http://dx.doi.org/10.1016/j.xpro.2020.100171 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Wang, Ying
Ma, Zhe
Damania, Blossom
Liu, Pengda
Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice
title Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice
title_full Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice
title_fullStr Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice
title_full_unstemmed Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice
title_short Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice
title_sort protocol for monitoring dna-triggered cgas/sting signaling in mammalian cells and mice
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757363/
https://www.ncbi.nlm.nih.gov/pubmed/33377065
http://dx.doi.org/10.1016/j.xpro.2020.100171
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