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. 2023 Dec 12;14:8217. doi: 10.1038/s41467-023-43001-y

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© The Author(s) 2023

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/.

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Fig. 1 — a, b Effects of cGAS overexpression on L1 retrotransposition efficiency in HeLa cells. n = 6 independent experiments for the 0.75 μg group, and n = 9 for the others. c, d Analysis of L1 retrotransposition efficiency in cGAS-knockout HeLa cells. n = 6 independent experiments. e, f Restoring cGAS in cGAS-deficient cells abolished the stimulatory effect of knocking out cGAS on L1 retrotransposition. n = 9 independent experiments. g Analysis of endogenous L1 copy number in cGAS-knockout cells. n = 3 independent experiments. h, i Analysis of endogenous L1 copy number in the kidney and brain of Cgas knockout mice (9 replicates from three independent mice per group). j, k Analysis of the protein level of ORF2p in cGAS-depleted HeLa cells. l Analysis of ORF2p protein levels in HeLa cells transfected with cGAS-HA. m Relative ORF2p abundance between the heavy and light isotope labeling groups assayed by SILAC-combined mass spectrometry. n, o Immunofluorescence analysis of ORF2p-induced γH2AX foci with or without cGAS overexpression in HeLa cells. The cells were transfected with a control vector or cGAS-HA vector together with an ORF2p-Flag vector. More than 50 cells were included for the analysis of each group (scale bar: 5 µm). p, q The effect of cGAS overexpression on genome stability in ORF2p-overexpressing HeLa cells. At least 50 cells per group were analyzed. r, s Immunofluorescence analysis of ORF2p-induced γH2AX foci in cGAS-knockout HeLa cells. More than 50 cells were included for the analysis of each group (scale bar: 5 µm). t, u The effect of knocking out cGAS on genome stability in ORF2p-overexpressing HeLa cells. At least 50 cells per group were analyzed. Data are presented as mean values ± s.d. for (a), (c), (e) and (g), and as mean values ± s.e.m. for (h) and (i). Student’s t test for (a), (c), (g), Brown-Forsythe and Welch ANOVA followed by Holm-Sidak’s multiple comparisons for (e), and the Mann–Whitney U test for (h), (i), (o), (q), (s) and (u). All statistical test used were two-sided. All experiments were repeated three times with similar results. Elements created with BioRender.com.