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. 2020 Aug 24;11:366. doi: 10.1186/s13287-020-01883-5

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Generation of reporter knock-in CRXp-tdTomato cell line. a Schematic diagram showing the targeting strategy of the insertion site. tdTomato cDNA sequence was fused in-frame into CRX behind start codon. b Overall 3D organoids fluorescent and bright field images on D45, D60, D90, and D120. A typical fluorescence intensity increasement along with differentiation time is presented. c Representative flow cytometry analysis in D45, D60, D90, and D120 organoids. Black and red represent the organoids derived from control and knock cell line, respectively. d Overall fluorescence intensity of organoids quantified by ImageJ, data are presented as the mean ± SEM (n = 27, 14, 11, and 10, respectively). e Proportion of tdTomato-positive cells in 3D organoids counted by flow cytometry, data are presented as the mean ± SEM (n = 6, 6, 8, and 3, respectively). f Comparison of increasement tendency between flow cytometry analysis and fluorescence intensity. g Relationship between fluorescence intensity and the number of tdTomato-positive cell in 3D organoids