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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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Observing rod production in live NRL^+/eGFP OVs. Phase and epifluorescence images of live NRL^+/eGFP OVs were obtained over time in live cultures. (A,B) Small patches of NRL-eGFP were detected beginning at d68 (arrowhead in B). (C–F) NRL-eGFP fluorescence increased from d78 (C,D) to d89 (E,F), by which time all OVs demonstrated robust NRL-eGFP fluorescence. (G,H) NRL-eGFP fluorescence is maintained to at least d230, the last time point examined. (G,I) Hair-like outer segment projections are formed along the outside of OVs (arrowheads). (J) To quantify the number of NRL-eGFP+ and NRL+ cells, dissociated OVs were plated and immunolabeled with an anti-NRL antibody. The number of NRL-eGFP+ and NRL+ cells increased dramatically and concomitantly from d70 to d95. At d130, roughly half of all cells within OVs were immunopositive for NRL-eGFP and NRL. Scale bar in H = 100 µm (applies to A–H); I = 50 µm.