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. 2022 Aug 16;13:4822. doi: 10.1038/s41467-022-32401-1

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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. 5 — a Reporter assays for IFNB1 promoter activity in HEK293T cells transfected with the indicated plasmids for 24 h. Data are presented as mean ± AD, n = 2 technical repeats. Data were analyzed by unpaired two-tailed Student’s t-test. b Interactions of STING with IRF8 and its mutants. HEK293T cells were transfected with the indicated plasmids for 20 h followed by co-immunoprecipitation and immunoblotting analysis with the indicated antibodies. EV, empty vector. c Effects of cGAMP stimulation on association of Sting with Irf8 or Irf8^S151A. HEK293 cells were transfected with the indicated plasmids for 24 h and then untreated or treated with cGAMP for 4 h. Co-immunoprecipitation and immunoblotting analysis were performed with the indicated antibodies. d ELISA analysis of Ifn-β and IP-10 secretion in WT and Irf8^−/− BMDMs reconstituted with the indicated plasmids. Cells were left uninfected or infected with HSV-1 for the indicated times. Data are presented as mean. n = 2 technical repeats. Data were analyzed by unpaired two-tailed Student’s t-test. e Detection of Irf8 phosphorylation in Irf8^−/− BMDMs reconstituted with Irf8 or Irf8^S151A. Cells were left uninfected or infected with HSV-1 for the indicated times followed by coimmunoprecipitation and immunoblotting analysis with the indicated antibodies. f Effects of S151 mutation on the association of its N-terminal region and IAD. The experiments were performed similarly as in b. g HSV-1-induced phosphorylation of Irf8^S151, Irf3^S388 and Tbk1^S172 in WT and Tbk1-knockout BMDMs. Cells were left uninfected or infected with HSV-1 for the indicated times. Immunoblotting analysis was performed with the indicated antibodies. h The Tbk1 inhibitor BX795 does not affect HSV-1-induced phosphorylation of Irf8^S151. BMDMs were untreated or treated with BX795 (1 μM) and then infected with HSV-1 for the indicated times. Immunoblotting analysis was performed with the indicated antibodies. Source data are provided as a Source data file.