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. Author manuscript; available in PMC: 2022 Aug 27.
Published in final edited form as: Science. 2018 Jul 5;361(6403):704–709. doi: 10.1126/science.aat1022

Fig. 3. Multivalent interactions drive cGAS–DNA condensation and promote cGAS activation.

Fig. 3

(A) Schematic of hypothetical cGAS and DNA valencies. (B) Representative images of phase separation by mixing cGAS (10 μM) with dsDNA of different lengths (10 μM) in physiological buffer. Scale bar: 10 μm. AF488: Alexa Fluor 488. (C) Bright-field images of phase separation by mixing DNA of different lengths with full-length or N-terminally truncated human or mouse cGAS as indicated. Scale bar: 20 μm. The images shown in (B) and (C) are representative of all fields in the wells. (D & E) cGAMP production by different concentrations of recombinant full length or ΔN human cGAS in low-salt buffer (D) or physiological buffer (E). Shown are the mean ± SD. N = 3 assays. (F) Quantification of cGAS–DNA puncta by imaging of MEF cells expressing GFP-tagged full length human cGAS or ΔN160-cGAS after transfection of Cy5-ISD. Representative images are shown in Figure S6. Values shown are means ± SD. N = 5 images. (G) cGAMP production in the MEF cells expressing full length or ΔN160 human cGAS after transfection with ISD or HT-DNA. Values are means ± SD. N = 3. Multiple t-tests; p-values (for F and G): > 0.0332 (n.s.), 0.0332 (*), 0.0021 (**), 0.0002 (***), < 0.0001 (****). cGAS expression levels are shown in Figure S5F. Data are representative of at least three independent experiments.