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. 2021 Aug 20;4:993. doi: 10.1038/s42003-021-02502-6

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© The Author(s) 2021

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. 2 — a–c IHC for Ki67 in mammary gland from lambs < 2 mo (a) and 5–9.5 mo (b) and accompanying mask derived using an algorithm detecting intra-epithelial Ki67 positive events (c). d Scatter plot demonstrating that the distributions of epithelial nuclear Ki67 positivity differ significantly between younger and older lambs. Dots represent individual lambs. Bars represent mean ± standard deviation. * p < 0.05. e, f IF for Ki67 (gold), α-SMA (cyan) and DNA (DAPI; magenta) demonstrating that the majority of Ki67 positive nuclei are in the luminal epithelial layer (arrowheads), with rare Ki67 positive nuclei in myoepithelial cells (arrow). e 1 do lamb. f 9.5 mo lamb. do, days old; mo, months old. Images are representative of eight (a), four (b), twelve (c) and three (e, f) biological repeats. All IHC images have haematoxylin counterstain. Scale bar = 200 μm (a–c); 100 μm (e); 50 μm (f).