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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2020
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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). |
format | Online Article Text |
id | pubmed-7757363 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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