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. 2021 Nov 11;12:6520. doi: 10.1038/s41467-021-26777-9

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© The Author(s) 2021

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. 4 — Representative flow cytometry plots and percentage of pHRodo⁺ airway macrophages (AM), depicting a efferocytosis and b phagocytosis capacities. c Gene expression of Nos2, Il6, Il1b, Il12b, and Mmp12, assessed by qPCR and d protein expression of IL-1α, IL-6, IL-23p40/p19, CCL2, and CXCL1, assessed by cytokine bead array, at indicated time points post lipopolysaccharide (LPS) treatment. PCA of e RNA sequencing (RNAseq) and f Assay for transposase-accessible chromatin sequencing (ATACseq) data from Scnn1b-transgenic (Tg) vs. wild-type (WT) AMs treated with lipopolysaccharide (LPS) for 12 h (h). g Functional enrichment analysis of the top 100 genes explaining PC1 (left panel) and PC2 (right panel). h Volcano plot visualizing the treatment response (LPS vs. medium) on chromatin accessibility level. differentially accessible regions (DARs): adjusted (adj.) P value < 0.05; absolute log2 fold change >2. Red dots: increased accessibility in LPS treated AMs; blue dots reduced accessibility in LPS treated AMs. i Profile plot of all identified LPS responsive DARs, visualized for LPS treated Scnn1-Tg and WT AMs (top panel) and primary uncultured Scnn1-Tg and WT AMs at baseline (lower panel). Box plots indicate the largest value within the 1.5× interquartile range above 75th percentile, 75th percentile, median, 25th percentile, and smallest value within the 1.5× interquartile range below 25th percentile of a n = 17 WT, n = 19 Scnn1b-Tg, and b n = 10 per group. *P value < 0.05; **P value < 0.01; ***P value < 0.001 per group by Mann–Whitney U-test. a P < 0.001, b P = 0.0011. Bar plots show mean ± SEM of c n = 10 per group (6, 12 h) and n = 9 per group (24 h). *P value < 0.05; **P value < 0.01; ***P value < 0.001 per group by Mann–Whitney U-test. Nos2: 6 h P = 0.0021, 12 h P < 0.001, 24 h P = 0.178; Il6: 12 h P = 0.0052, 24 h P = 0.0078; Il1b: 12 h P = 0.0048; Il12b: 12 h P = 0.0089, 24 h P = 0.0314; Mmp12: 6 h P = 0.0021, 12 h P = 0.0887, 24 h P < 0.001. d Lipopolysaccharide (LPS) treatment: IL-1α, IL-23p40/p19, IL-6, CXCL1: n = 10 per group (6, 12 h), n = 9 per group (24 h); CCL2: n = 10 wt (6, 12 h), n = 9 wt (24 h), n = 9 Scnn1b-Tg (6, 12, 24 h). Medium control: IL-1α, IL-23p40/19, CCL2, CXCL1: n = 8 WT (6 h), n = 10 WT (12 h), n = 9 WT (24 h), n = 9 Scnn1b-Tg (6, 12, 24 h), IL-6: n = 9 WT (6 h), n = 10 WT (12 h), n = 9 WT (24 h), n = 9 Scnn1b-Tg (6, 12, 24 h). *P value < 0.05; **P value < 0.01; ***P value < 0.001 by One-Way ANOVA followed by Tukey´s post hoc test. e–i n = 3 per group. ND not detectable. Source data are provided as a Source Data file.