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. 2017 Aug 15;7:8295. doi: 10.1038/s41598-017-07603-z

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© The Author(s) 2017

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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Temporal delay in Zpr1 mutation reduces severity of defects in the development of respiratory system. Whole diaphragms from E18.5 embryos were stained with neurofilament (NF) antibody. Images shown are composite of individual panels acquired by TileScan using confocal microscope. Diaphragms from (a,a’) control (b,b’) Zpr1 ^Hb9MNΔ and (c,c’) Zpr1 ^ChATMNΔ mice showing innervation of phrenic nerve in the muscle. Comparison of the phrenic nerve staining between Zpr1 ^Hb9MNΔ and Zpr1 ^ChATMNΔ mice shows remarkable improvement in the development and innervation of phrenic nerve upon temporal delay in Zpr1 mutation in motor neurons. Arrowheads indicate phrenic nerve. Scale bar is 1000 μm.