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. 2020 Nov 26;11:6000. doi: 10.1038/s41467-020-19849-9

Fig. 4. β-arrestin 2 interacts with cGAS and promotes the activation of cGAS.

Fig. 4

a Volcano plots comparing gene expression in wild-type versus Arrb2−/− infected peritoneal macrophages. b GO enrichment analysis of differential genes involved in the terms as indicated. c KEGG enrichment analysis of differential genes involved in the pathways as indicated. d, e Luciferase assay of IFN-β d (***P < 0.0001, **P = 0.0033, 0.0021 in sequence, ***P < 0.0001) and ISRE e (**P = 0.0052, ***P = 0.0001, **P = 0.0010, 0.0012 in sequence) activation in HEK293T cells expressing various vectors. f, g Immunoassay of lysates of HEK293T cells expressing various vectors. h Direct binding of His–β-arrestin 2 with GST-cGAS. i, j Luciferase assay of IFN-β i (***P < 0.0001, <0.0001 in sequence, **P = 0.0003) and ISRE j (**P = 0.0001, ***P < 0.0001, < 0.0001 in sequence) activation in HEK293T cells expressing various vectors. k Immunoassay of cell lysates and streptavidin-precipitated proteins from HEK293T cells transfected with various vectors and stimulated with biotin-ISD. l EMSA of cGAS and dsDNA. m Immunoassay of lysates of HEK293T cells expressing various vectors. n cGAMP activity measured by extracts from HT-DNA-stimulated (6 h) WT and Arrb2−/− peritoneal macrophages. o–q Immunoblot analysis of monomeric and dimeric STING in WT and Arrb2−/− peritoneal macrophages o, control siRNA or siβ-arrestin 2-transfected PAW264.7 cells p, and control or β-arrestin 2 vector-transfected L929 cells q. Data are representative of at least three independent experiments (mean ± SEM in d, e, i, j, n = 3). Two-tailed unpaired Student’s t-test.