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. 2018 Feb 5;8:2370. doi: 10.1038/s41598-018-20813-3

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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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eGFP labeling of rods in mid-stage NRL^+/eGFP OVs. To examine mid-stages of rod differentiation in NRL^+/eGFP OVs, d145 OVs were analyzed. (A,B) A large increase in rod production was observed by d145, as shown by co-expression of NRL-eGFP+ (A) and NR2E3 (B). (C,D) Numerous ARR3+ cones were also present at this time (C; merged image in D). (E) Higher magnification image of showing non-overlapping NRL-eGFP and ARR3 labeling in rods and cones. (F–H) Many NRL-eGFP+ rods (F) also expressed rhodopsin (RHO) (G; merged image in H) at this stage of differentiation. (I–P) Analysis of NRL-eGFP and NRL expression relative to red/green cone opsin (OPN1MW/LW; I–L) or blue cone opsin (OPN1SW; M–P) expression showed complete segregation of all cone opsins from NRL-eGFP+/NRL+ cells. Note the predominant localization of cone opsin-expressing cells along the outer (apical) portion of OVs (asterisks in K), although some cones were also found beneath the organized photoreceptor outer nuclear layer (arrowhead in K). Scale bars = 50 µm.