Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Jan 10;625(7995):585–592. doi: 10.1038/s41586-023-06889-6

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Extended Data Fig. 7 — a, b, c, d, Electrophoretic mobility shift assays (EMSA) a, Native gel of nucleosomes containing 147 or 185 bp DNA in presence of increasing concentrations of MRN complex, stained with ethidium bromide. b, Native gel of 147 bp nucleosomes containing either wild type or acidic patch mutant (APM) histones in the presence of increasing concentrations of MRN complex, stained with ethidium bromide. c, Native gel as in panel (b) using 185 bp nucleosomes with 20 bp symmetric linker on each side of the nucleosome core. d and e, Native gel of 50 nM (d) or 100 nM (e) 147-bp hNCPs without or with 200 nM (d) or 50 nM (e) carboxyrhodamine-labelled mouse cGAS catalytic domain (mcGAScat-CR) and 0-1.2 μM MRN. The upper gel shows ethidium bromide staining of DNA, and the lower gel shows fluorescence signal from mcGAScat-CR. hNCP alone as well as 1:1 and 2:1 cGAS:hNCP complexes are indicated. (The results from the tests (a, b, c, d, e) demonstrate consistent findings across three independent replicates. f, Quantification of bands from the corresponding gels (e). g, TR-FRET assay for detection of nucleosome stacking mediated by human cGAS catalytic domain (hcGAScat) or MRN complex. EU-SA, LANCE Eu-W1024 Streptavidin; AT647N, Atto647N; biotin-NUC, H2BK125C-biotin 147-bp nucleosomes. Data are mean ± s.d. h, i, Substrate, intermediate, and product are ATP, pppGpA, and cGAMP, respectively. h, 1 μM cGAS: 5 μM dsDNA, 0.5 μM hNCP, 0–2 μM MnucdeadRN (containing Mre11^H129N nuclease deficient protein), demonstrating consistent findings across three independent replicates. i, 0.5 μM cGAS: 5 μM dsDNA, 0.5 μM hNCP, 0–2 μM MRN demonstrating consistent findings across two independent replicates. j, Quantification of standardized percentage of cGAMP product across three biological replicates demonstrates greater cGAS activity in the presence of increasing concentrations of MRN, but only when the cGAS:hNCP ratio is 2:1 versus 1:1. Shown are the mean value with error bars representing the standard deviation.

Source Data