Lineberger 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.
STAR Protoc. 2020 Nov 14;1(3):100171. doi: 10.1016/j.xpro.2020.100171. eCollection 2020 Dec 18.
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).
先天免疫是哺乳动物抵御感染的第一道防线，其受到严格调控。我们通过 Western blot、RT-PCR 和 ELISA 分析监测了 ISD90 或 2',3'-cGAMP 刺激 EA.hy926 细胞后 cGAS/STING 信号的变化，以揭示信号的激活和 IFNβ 的产生。此外，我们还包括 HSV-1 感染的小鼠模型，以进一步揭示在小鼠中分析 cGAS/STING 信号的步骤。该方案可应用于专注于细胞培养或小鼠模型的研究，以研究 cGAS/STING 信号。有关该方案使用和执行的完整详细信息，请参阅 Zhang 等人。（2020 年）。
