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

Highlights

• A protocol to monitor cGAS/STING signaling at protein and mRNA levels

• Western blotting analyses to monitor key phosphorylation events

• RT-PCR and ELISA analyses to determine IFNβ levels

• Mouse model of HSV-1 infection to measure cGAS/STING in vivo

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).