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. 2022 May 10;5:433. doi: 10.1038/s42003-022-03400-1

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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 The cGAS KO(LL/RK-NLS) RAW264.7 cells were treated with 2 μg/mL Dox for 24 h, followed by mock-infection or infection with 1 MOI of HSV-1 McKrae for 16 h. Then, cells were fractionated into five fractions and blotted as indicated. STING: membrane marker; TBK1 and α-tubulin: cytosolic marker; H3: nuclear marker. Red arrow indicates nuclear soluble cGAS. b, c The cGAS KO, KO(mcGAS), KO(LL/RK-NLS) RAW264.7 cells were treated with Dox for 24 h and then infected with 1 MOI of HSV-1 KOS d109 for designated times. Real-time PCR was performed to determine the relative mRNA levels of IFNβ (b) and IP-10 (c). Data represent means ± s.d. of three independent experiments. The P-value was calculated by two-way ANOVA followed by Dunnett’s multiple comparisons test. d The cGAS KO, KO(mcGAS), KO(LL/RK-NLS) RAW264.7 macrophages were treated with Dox for 24 h and then infected with 1 MOI of HSV-1 KOS d109 for indicated times. Cell lysates were collected and blotted as indicated. e cGAS KO, KO(mcGAS), KO(LL/RK-NLS) RAW264.7 macrophages were treated with Dox for 24 h and then infected with 1 MOI of HSV-1 KOS d109 for 8 h. The amount of cGAMP in each cell line was determined by ELISA assays. All experiments were repeated three times. The P value was calculated by one-way ANOVA followed by Tukey’s multiple comparisons test. f The cGAS KO(LL/RK-NLS) RAW264.7 cells were treated with DMSO or 25 μM RU.521 for 16 h. Then, cells were mock-infected or infected with HSV-1 KOS d109 for 16 h. Real-time PCR was performed to determine the relative mRNA levels of IFNβ, IP-10, and RANTES. Data represent means ± s.d. of three independent experiments. The P-value was calculated by two-way ANOVA followed by Tukey’s multiple comparisons test. g The cGAS KO(LL/RK-NLS) and KO(LL/RK-NLS)/D307A cells were treated with Dox for 24 h and then mock-infected or infected with HSV-1 KOS d109 for 16 h. Real-time PCR was performed to determine the relative mRNA levels of IFNβ, IP-10, and RANTES. Data represent means ± s.d. of three independent experiments. The P-value was calculated by two-way ANOVA followed by Sidak’s multiple comparisons test.