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. 2022 Oct 10;13:5973. doi: 10.1038/s41467-022-33671-5

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a Denature-IP (with anti-FLAG) and immunoblot analysis (with anti-FLAG, anti-HA, anti-GFP, or anti-ISG15) of HEK293 cells transfected with plasmids encoding FLAG-cGAS, HA-UBE1L, His-ISG15, and GFP-UBCH8 together with an empty vector, HA-ARIH1 or HA-ARIH1^C357S for 24 h. b GST-cGAS was incubated with UBE1L, UBCH8, His-ISG15, and His-ARIH1, or His-ARIH1^C357S in the reaction buffer for 2 h and subject to 13.5% SDS-PAGE followed by Coomassie brilliant blue staining. c Denature-IP (with anti-cGAS) and immunoblot analysis (with anti-cGAS, anti-ARIH1, anti-GAPDH, or anti-ISG15) of Cre-ER and Cre-ER;Arih1^fl/fl MLFs that were treated with 4-OHT (1 μM) for 3 days followed by HSV-1 infection for 0–6 h. d Denatire-IP (with anti-cGAS) and immunoblot analysis (with anti-cGAS, anti-ISG15, anti-ARIH1, or anti-Tubulin) with SDS-PAGE or SDD-AGE of 4OHT (1 μM, 3 days)-treated Cre-ER and Cre-ER;Arih1^fl/fl MLFs that were reconstituted with different amounts of Lenti-FLAG-ARIH1 followed by infection with HSV-1 for 6 h. e cGAMP activity assay of cells treated as in (d) that were transfected with HSV120 (1 μg) for 6 h. f Denature-IP (with anti-FLAG) and immunoblot analysis (with anti-FLAG, anti-HA, anti-GFP, or anti-ISG15) of HEK293 cells transfected with plasmids encoding FLAG-cGAS, FLAG-cGAS^K187R, HA-UBE1L, His-ISG15, and GFP-UBCH8 together with an empty vector, HA-ARIH1 or HA-ARIH1^C357S for 24 h. g mcGAS or mcGAS^K173N was incubated with UBE1L, UBCH8, His-ISG15, and GST-ARIH1 in the reaction buffer for 2 h and subject to gradient SDS-PAGE followed by Coomassie brilliant blue staining. h Denature-IP (with anti-FLAG) and immunoblot analysis (with anti-FLAG, anti-ARIH1, anti-GAPDH, or anti-ISG15) of cGas^−/− MEFs reconstituted with FLAG-mcGAS or mcGAS^K173N followed by HSV-1 infection for 0–6 h. Data are representative of three independent experiments (a–h) Graphs show mean ± S.D. (n = 4 for (e), biologically independent experiments). Statistical significance was determined by one-way ANOVA in (e) Source data are provided as a Source Data file.