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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. 1 — a Yeast-two hybrid assays with AH109 cells that were co-transformed with pGBT9-cGAS and pGADT7-E3 constructs. PC positive control (pGBT9-MAVS and pGAD7-RNF115); NC negative control (pGBT9-cGAS and pGADT7 empty vector). b Immunoprecipitation (with anti-FLAG) and immunoblot analysis (with anti-FLAG or anti-HA) in HEK293 cells that were transfected with plasmids encoding FLAG-ARIH1 and HA-tagged, RIG-I, MAVS, cGAS, MITA, TBK1 or IRF3 for 24 h. c Immunoprecipitation (with control IgG or anti-cGAS) and immunoblot analysis (with anti-cGAS, anti-ARIH1, or anti-GAPDH) in MLFs (left) or BMDCs (right) that were left uninfected or infected with HSV-1 for 4–8 h. d Immunoblot analysis (with anti-FLAG) of the GST-pulldown precipitants from the incubation of GST or GST-ARIH1 (5 μg) and in vitro translated FLAG-cGAS. The input GST and GST-ARIH1 were subject to SDS-PAGE followed by Coomassie brilliant blue staining. e, f Immunoprecipitation (with anti-FLAG) and immunoblot analysis (with anti-FLAG or anti-HA) in HEK293 cells that were transfected with plasmids encoding HA-cGAS and FLAG-tagged ARIH1 or truncates (e) or with plasmids encoding FLAG-ARIH1 and HA-tagged cGAS or mutants (f) for 24 h. UBA Ubiquitin-associated domain, RING1 RING domain 1, IBR in-between RING domain, RING2 RING domain 2, AD Ariadne domain, NTD N-terminal domain, NTase nucleotidyl transferase, Mab21 Mab-21 domain. Data are representative of three independent experiments (a–f). Source data are provided as a Source Data file.