May 30, 2013 · Then, the resulting cGAS products were purified and transfected into HEK 293T cells expressing STING variants. In contrast to synthetic cGAMP, the cGAS product was able to activate hSTINGREF and mSTING R231A (Figure 2 C).

Apr 8, 2021 · Intracellular 2′3′-cGAMP concentrations determined STING translocation or condensation. STING biocondensates constrained STING and TBK1 (TANK binding protein 1) to prevent innate immunity...

In this study, we report that SV40 LT inhibits cGAS-STING interferon induction through a mechanism distinct from other viral oncogenes and uncover an unanticipated defect underlying the loss of STING in 293T cells.

Feb 4, 2023 · To verify that the construct was functional, we performed microscopy in 293T cells stably expressing STING-TurboID and confirmed STING translocation upon cGAMP stimulation and co-localization...

Immunoprecipitated cGAS from 293T cells transiently expressing FLAG-tagged human cGAS was resolved by SDS-PAGE and analyzed by mass spectrometry. We found that human cGAS was phosphorylated at S305 when overexpressed in 293T cells (Figure S1 A).

Jan 24, 2020 · We show that STING transcription is suppressed in 293T cells but is not driven by SV40. This study also revealed that SV40 LT does indeed inhibit cGAS-STING interferon induction, but through a mechanism distinct from other DNA virus oncogenes.

In ENPP1 knockout 293T cells, a significant increase of extracellular cGAMP molecules was detected (92), demonstrating the potential of ENPP1 in regulating the cGAS-STING signaling.

On the contrary, we detected much lower extracellular cGAMP levels in 293T cells, which have very low cGAS expression, and in primary human PBMCs, which have high cGAS expression, but likely no cytosolic dsDNA to stimulate cGAS activity (Fig. 4b).

To test if cGAS binds to the nucleosome directly, we purified nucleosomes from HEK 293T cells and reconstituted nucleosomes using histone proteins 20 (Extended Data Fig. 1a, b). Next, we conducted cGAS binding studies using mouse cGAS (mcGAS) catalytic domain.

Mar 21, 2017 · Here we report that caspases controlled antiviral immunity through cGAS cleavage during inflammasome activation. Upon inflammasome activation, caspase-1 directly bound to and cleaved human cGAS at D 140/157, resulting in reduced …