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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. 3 — Neutrophil extracellular traps (NETs) induced by cigarette smoke extract (CSE-NETs) dose-dependently promote the proliferation and the gene expressions of both nuclear factor kappa B (NF-κB)-dependent inflammatory cytokines and type-I interferons (IFNs) in the human airway epithelial cells (hAECs); CSE-NETs promote the maturation of human dendritic cells (DCs). Statistical analysis: n = 3–16 for each bar in (a–m) from at least three independent experiments, data were presented as the mean ± standard deviation; Differences are assessed by the (a–l) one-way or (m) 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. a–c Human peripheral neutrophils were stimulated by 5% CSE for 18 h to induce NETs, which were subsequently collected and quantified for DNA concentrations. Effects of 6, 12, and 24 μg/mL NETs (incubated for 48 h) on the proliferation of hAECs, as assessed by the a EdU proliferation assay (supplementary Method 11) and mRNA expressions of b MKI67 and c PCNA (both are markers of proliferation, supplementary Method 13). d–l Effects of 6, 12, and 24 μg/mL NETs on the mRNA expressions and soluble levels of NF-κB-dependent inflammatory cytokines and type-I IFNs of hAECs (supplementary Method 13, 26): d mRNA expression of CXCL5 (C-X-C motif chemokine ligand 5), e mRNA expression of CXCL8, f mRNA expression of TNFα (tumour necrosis factor-alpha), g mRNA expression of IL-1β (interleukin 1β), h soluble levels of IL-1β in cell-culture supernatants, i soluble levels of CXCL8 in cell-culture supernatants, j mRNA expression of IFN-β1, k mRNA expression of IL-12 and l soluble levels of IFN-β in cell-culture supernatants. m, n 12 μg/mL of NETs promote the maturation of dendritic cells (differentiated from peripheral blood monocytes) from both healthy participants and patients with COPD (supplementary Method 15); the maturation is evaluated by m the percentage of CD86⁺ CD40⁺ cells, as assessed by flow cytometry with a gating strategy illustrated in (n) (Supplementary Method 17)