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. 2020 Aug 12;11(8):683. doi: 10.1038/s41419-020-02848-5

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© The Author(s) 2020

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. 3 — Adenovirus-expressing GFP-PAX6 (PAX6 group) or GFP vector (Control group) was separately injected into the anterior chamber of rat eyes. a GFP expression of corneal endothelium indicated the transfection efficiency of GFP-encoding vector and PAX6 adenovirus. White arrows showed that representative GFP⁺ cells had increased the level of PAX6 protein in the PAX6 group, but not the Control group. The dotted lines highlighted the boundary between the endothelium (En) and the anterior segment. b–d PAX6 protein was overexpressed in corneal endothelium of rats via adenovirus injection through the anterior chamber. Then, the rat corneal endothelial injury model was induced by cryoinjury 3 days after injection. b, c Representative images of slit-lamp microscope (b) and optical coherence tomography (OCT) (c) showed opacity and edema of corneas at 24, 48, and 72 h after injury. Note: Corneal opacity and corneal edema were lower in the PAX6 group, compared with the Control group. d Analysis of central corneal thickness in control and PAX6 groups at all investigated time points. e, f Normal and injured corneas were acquired for organ culture. GFP-vector and GFP-PAX6 adenovirus-transfected CECs in vitro and corneas were collected for ZO-1 staining. e Representative flat-mounted images of ZO-1 expression in the normal CECs. f ZO-1 staining of injured CECs from the Control and PAX6 groups. Note that PAX6 overexpression promoted tight junction among CECs after injury. Scale bar = 20μm. Error bars showed means ± SEM (n = 3/group; *p < 0.05, **p < 0.01).