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. 2019 Aug 2;9:11277. doi: 10.1038/s41598-019-47653-z

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© The Author(s) 2019

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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Collagen distributions within the developing mouse cornea. All corneal epithelia are at the top in E14 and E16, and to the right in E12 and E18. Type I collagen was initially seen within the surface ectoderm (SE) (blue arrow) and lens capsule (LC) (yellow arrow) at E12, with enhanced expression posterior to the corneal epithelium (CE) at E14 (blue arrows) and within the anterior corneal stroma (red arrows). Expression was shown throughout the corneal stroma from E16-E18 (red arrows). Type I collagen was enhanced posteriorly to the corneal epithelium at E16 (blue arrows) as well as anterior to the endothelium at E16 and E18 (green arrows). Type II collagen was initially seen within the surface ectoderm (blue arrow) and lens capsule (yellow arrow) at E12, enhancing within the anterior cornea from E14 and throughout the corneal stroma between E16-E18 (red arrow). Expression was also increased posterior to the corneal epithelium at E16 (blue arrow). Type V collagen was expressed within the surface ectoderm as a localised line (blue arrow) as well as within the lens capsule (yellow arrow) from E12, type V collagen expressed posteriorly to the corneal epithelium from E14 (blue arrow) and throughout the corneal stroma from E16-E18 (red arrows). An enhanced expression was also seen posteriorly to the corneal epithelium from E16-E18 (blue arrow) and anteriorly to the endothelium from E18 (green arrow). Type IX collagen was not seen at E12, with a little expression associated with the corneal epithelium at E14 (blue arrows). Expression enhanced between E16-E18 within the corneal epithelium (blue arrows). No type IX collagen was seen within other areas of the cornea during development. No expression was apparent within the no primary antibody control and the rabbit IgG control images (data not shown). Adult corneas were used as a positive control for all antibodies (data not shown). Blue (DAPI – cell nucleus), red (collagen). Scale bar = 50 μm.