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

Summary: 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....

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Main Authors:	Ying Wang, Zhe Ma, Blossom Damania, Pengda Liu
Format:	Article
Language:	English
Published:	Elsevier 2020-12-01
Series:	STAR Protocols
Subjects:	Antibody Cell culture Cell-based Assays Immunology Signal Transduction
Online Access:	http://www.sciencedirect.com/science/article/pii/S2666166720301581

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spelling	doaj-7b02a563d739469786aceef909c5fb152020-12-21T04:48:59ZengElsevierSTAR Protocols2666-16672020-12-0113100171Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and MiceYing Wang0Zhe Ma1Blossom Damania2Pengda Liu3Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USADepartment of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610, USALineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; University of North Carolina Center for AIDS Research, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Corresponding authorLineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166720301581AntibodyCell cultureCell-based AssaysImmunologySignal Transduction
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Ying Wang Zhe Ma Blossom Damania Pengda Liu
spellingShingle	Ying Wang Zhe Ma Blossom Damania Pengda Liu Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice STAR Protocols Antibody Cell culture Cell-based Assays Immunology Signal Transduction
author_facet	Ying Wang Zhe Ma Blossom Damania Pengda Liu
author_sort	Ying Wang
title	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_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_sort	protocol for monitoring dna-triggered cgas/sting signaling in mammalian cells and mice
publisher	Elsevier
series	STAR Protocols
issn	2666-1667
publishDate	2020-12-01
description	Summary: 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).
topic	Antibody Cell culture Cell-based Assays Immunology Signal Transduction
url	http://www.sciencedirect.com/science/article/pii/S2666166720301581
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Protocol for Monitoring DNA-Triggered cGAS/STING Signaling in Mammalian Cells and Mice

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