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. 2022 Jun 21;13:3544. doi: 10.1038/s41467-022-31149-y

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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. 2 — a Expression profile of K⁺ channels in mouse bone marrow-derived macrophages (BMDMs) analyzed by RNA-seq (n = 3). The FPKM data of different genes are standardized to the Z-score by column in IBM SPSS Statistics. b–d Electrophysiological properties of freshly isolated WT (b–d) and Kir2.1-knockout (d) mouse peritoneal macrophages using patch-clamp. Thioglycolate-elicited mouse peritoneal macrophages were treated with (b) or without (c) 500 ng/ml LPS for 1 h before recording. The time-dependent current traces were recorded at a clamp voltage of –115 mV or +60 mV (red arrow, initiation of recording). The lower panel shows the I–V curves of the corresponding points for different doses of ML133 in the left panel. d Representative I–V curves of WT and Kir2.1-knockout mouse peritoneal macrophages. e Statistics of current amplitude recorded at a clamp voltage of –115 mV. Macrophages were treated and clamped as described above (Kcnj2^f/f, n = 16, 18, 8, and 17 respectively; Lyz2-cre-Kcnj2^f/f, n = 16, 9, 10, and 10 respectively; mean ± SEM). f Statistics of Vm recorded at 3 min after initiation of recording (Kcnj2^f/f, n = 10, 7, 8, 9, 9, 8, 8, and 7, respectively; Lyz2-cre-Kcnj2^f/f, n = 9, 8, 12, and 12, respectively; mean ± SEM). g, h RNA-seq analysis of inflammatory genes. GSEA analysis showing enrichment of the inflammatory response genes (g) and heatmap of inflammatory genes (h). NES, normalized enrichment score; FDR, false discovery rate. (n = 3). i Il1a, Il1b, and Tnf mRNA transcription assays. Mouse peritoneal macrophages from WT or Lyz2-cre-Kcnj2^f/f mice treated with 500 ng/ml LPS for 6 h in the presence or absence of ML133 (25 μM) followed by qPCR analysis (n = 6, mean ± SEM). j Mouse peritoneal macrophages treated with or without 500 ng/ml LPS in the presence or absence of elevated [K⁺]_e (50 mM), gramicidin (1.25 μM), ML133 (25 μM), PAP1 (Kv1.3 inhibitor, 1 μM), Tram34 (KCa3.1 inhibitor, 1 μM) for 6 h followed by qPCR analysis of mRNA transcription. Two-tailed unpaired Student’s t-test. Source data are provided as a Source Data file.