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. 2024 Jun 17;9:163. doi: 10.1038/s41392-024-01881-6

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© The Author(s) 2024

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. 4 — Cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS), and toll-like receptor 9 (TLR9) are required for the neutrophil extracellular traps (NETs)-stimulated proliferation, expressions of both nuclear factor kappa B (NF-κB)-dependent inflammatory cytokines and type-I interferons (IFNs) in human airway epithelial cells (hAECs), and maturation of human dendritic cells (hDCs). Statistical analysis: n = 6–10 for each bar in (a–x), n = 3 for each bar in y from at least three independent experiments, data were presented as the mean ± standard deviation; Differences are assessed by the a–y two-way ANOVA analysis of variance, followed Tukey’s honest significant test; P < 0.05 represents a significant difference, the scattered samples and the p values are displayed in figures. Effects of a–c cGAS and m–o TLR9 silencing on 12 μg/mL of NETs-stimulated proliferation of hAECs, as assessed by the a, m EdU proliferation assay (supplementary Method 11), and the mRNA expression of b, n MKI67 and c, o PCNA (both are markers of proliferation, supplementary Method 13). Effects of d–l cGAS and p, x TLR9 silencing on 12 μg/mL of NETs-induced mRNA expression and soluble levels of NF-κB-dependent inflammatory cytokines and type-I interferons (IFNs) in hAECs (supplementary Method 13, 26): d, p mRNA expression of CXCL5 (C-X-C motif chemokine ligand 5), e, q mRNA expression of CXCL8, f, r mRNA expression of TNFα (tumour necrosis factor-alpha), g, s mRNA expression of IL-1β (interleukin 1β), h, t soluble levels of IL-1β in cell-culture supernatants, i, u soluble levels of CXCL8 in cell-culture supernatants, j, v mRNA expression of IFN-β1, k, w mRNA expression of IL-12 and l, x soluble levels of IFN-β in cell-culture supernatants. y The inhibition of cGAS and TLR9 by 5 μM of RU.521 and 2 μM of ODN 2088, respectively, reduces 12 μg/mL of NETs-mediated maturation of hDCs (supplementary Method 16). z Representative flow cytometry images display the gating strategy and maturation of DCs treated with either RU.521 or ODN 2088, as evaluated by the percentage of CD86⁺ CD40⁺ cells (supplementary Method 17)