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

STAR Protoc. 2020 Nov 14;1(3):100171. doi: 10.1016/j.xpro.2020.100171. eCollection 2020 Dec 18.

Abstract

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).

Keywords: Antibody; Cell culture; Cell-based Assays; Immunology; Signal Transduction.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Assay / methods*
  • Cell Line
  • DNA / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Interferon-beta / metabolism
  • Mammals / metabolism*
  • Membrane Proteins / metabolism*
  • Mice, Inbred C57BL
  • Nucleotidyltransferases / metabolism*
  • Signal Transduction*

Substances

  • Membrane Proteins
  • Interferon-beta
  • DNA
  • Nucleotidyltransferases