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. 2018 Nov 2;9:4600. doi: 10.1038/s41467-018-06833-7

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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. 1 — The ENU-induced mutant exhibits lung development defects. a Gross morphology of wild-type (n = 15) and mutant (n = 12) P0 mice. b Floating assay for wild-type (n = 10) and mutant (n = 8) P0 lungs. c Representative pictures of distal lung from wild-type (n = 10) and mutant (n = 8) P0 mice. d Hematoxylin and eosin (H&E) staining and morphometric analysis of alveolar space and septal thickness in wild-type (n = 15) and mutant (n = 10) P0 lungs. e H&E staining of wild-type (n = 6 for each stage) and mutant (n = 5 for each stage) lungs at E16.5, E17.5, and E18.5. f PAS staining (n = 15 wild types, n = 10 mutants) and phenotype quantification at P0. g Representative TEM images of AT2 cells in wild-type (n = 3) and mutant (n = 3) lungs at E18.5. Arrowheads point to lamellar bodies in AT2 cells; g glycogen. h Immunostaining for PDPN in wild-type (n = 8 per each stage) and mutant (n = 8 per each stage) lungs at E18.5 and P0. Arrowheads point to AT1 nuclei; yellow dotted lines outline AT1 cell morphology. Error bars are means ± s.e.m. ****P < 0.0001, two-tailed Student’s t-test. Scale bars: 500 μm (c), 50 μm (d, e), 30 μm (f, h), 2 μm (g)