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. 2017 Oct 31;8:1220. doi: 10.1038/s41467-017-01332-7

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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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Fig. 6 — Cell density regulates RBC axon and synapse development. a–c Maximum intensity projection of RBC axon terminals labeled by transgenic expression of tdTomato (Grm6 _L -tdTomato in blue) and presynaptic release sites stained for the C-terminal binding protein 2 (CtBP2, red) in wild-type (a), Pcp2-DTA (b), and Pax6-DTA (c) retinas. For visual clarity, RBC axons and their synapses were digitally isolated in Amira. Scale bar indicates 5 μm. d–f Summary data of RBC axon surface areas in wild-type (d, n = 16 RBCs), Pcp2-DTA (e, n = 17 RBCs), and Pax6-DTA (f, n = 15 RBCs) mice. By Kruskal–Wallis one-way ANOVA testing, RBC axon surface areas in Pcp2-DTA and Pax6-DTA retinas were greater than in wild-type retinas (p < 0.02 and p < 10⁻⁸, respectively), and RBC axon surface areas were greater in Pax6-DTA than in Pcp2-DTA retinas (p < 0.003). g–i Population data (mean ± SEM) from wild-type (g, n = 16 RBCs), Pcp2-DTA (h, n = 17 RBCs), and Pax6-DTA (i, n = 15 RBCs) retinas show that synapse density tends to decrease with increasing axon size. By Kruskal–Wallis one-way ANOVA testing, RBC synapse densities in Pcp2-DTA and Pax6-DTA retinas were lower than in wild-type retinas (p < 0.003 and p < 0.02, respectively)