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. 2018 Jul 19;9:2815. doi: 10.1038/s41467-018-05043-5

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© The Author(s) 2018

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 — Kcnj13 regulates membrane potential and actin organization in SM cells. a DiBAC4(3) fluorescence in dorsal views of E14.5 WT (n = 9) and Kcnj13^T38C/T38C (n = 9) tracheal SM cells. b Quantification of mean DiBAC4(3) fluorescence intensity in E14.5 WT (n = 9) and Kcnj13^T38C/T38C (n = 9) tracheal SM cells. c DiBAC4(3) fluorescence in dorsal views of E14.5 tracheal SM cells after DMSO (n = 6) or 50 μM VU590 (n = 6) treatment. d Quantification of mean DiBAC4(3) fluorescence intensity in E14.5 tracheal SM cells after DMSO (n = 6) or 50 μM VU590 (n = 6) treatment. e Immunostaining for αSMA (red) and phalloidin (green), and DAPI (blue) staining in dorsal views of E14.5 WT (n = 8) and Kcnj13^T38C/T38C (n = 8) tracheas. f Quantification of mean phalloidin fluorescence intensity in E14.5 WT (n = 8) and Kcnj13^T38C/T38C (n = 8) tracheal SM cells. g Representative images of ventral views of E12.5 tracheas after a 48 h DMSO (n = 6) or 50 μM VU590 (n = 6) treatment. Double-sided arrows indicate tracheal tube length. h Quantification of relative tracheal tube length after a 48 h DMSO (n = 6) or 50 μM VU590 (n = 6) treatment. i Immunostaining for αSMA (red) and phalloidin (green), and DAPI (blue) staining in dorsal views of E12.5 tracheas after 48 h DMSO (n = 9) or 50 μM VU590 (n = 9) treatment. Quantification of E12.5 tracheal SM cell orientation (j), NAR (k), and mean phalloidin fluorescence intensity (l) after a 48 h DMSO (n = 9) or 50 μM VU590 (n = 9) treatment. m Immunostaining for αSMA (red) and DAPI staining (blue) in dorsal views of E12.5 WT (n = 24) and Kcnj13^T38C/T38C (n = 12) tracheas after a 48 h DMSO treatment, and Kcnj13^T38C/T38C tracheas (n = 12) after a 48 h 2 μM ionophore valinomycin treatment. Quantification of SM cell orientation (n) and NAR (o) of E12.5 WT (n = 12) and Kcnj13^T38C/T38C (n = 12) tracheas after a 48 h DMSO treatment, and Kcnj13^T38C/T38C tracheas (n = 12) after a 48 h 2 μM ionophore valinomycin treatment. Scale bars: 500 μm (e), 20 μm (a, c, g, i, m). *P < 0.05; **P < 0.01; unpaired Student’s t-test, mean ± s.d. Cont: control, Val: valinomycin