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. 2022 Mar 10;5:219. doi: 10.1038/s42003-022-03161-x

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© The Author(s) 2022

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 (left) Cluster-assigned bubble plot of cluster-specific DEGs encoding cell-surface markers confirms expression of CD200 and AMIGO2 in cluster 3 (arrows). (right, upper) Contour FACS plots of crude preparation of breast organoids isolated with Trop2 and CD271, followed by gating for MEP cells (encircled with the approximate percentage of cells indicated), subsequently sorted according to CD200 and CD271 to isolate CD200^low (blue) and CD200^high (red) cells, respectively. (right, lower) Bar graph of the fold change of normalized relative gene expression by RT-qPCR show upregulation of CD200, AMIGO2 and FAM126A, and downregulation of RARRES1, TXNIP and TNFSF10 in CD200^high (red) versus CD200^low (blue). Bars indicate mean ± SEM (***p < 0.005 by multiple t tests with Bonferroni correction, n = 3). b (left) Violin plots of KRT14 and KRT17 expression levels reveal upregulation of both in cluster 3. (right) Bar graph of quantification of immunostaining of smears from CD200^low (blue) and CD200^high (red) MEP cells confirms a significantly higher frequency (% K17⁺ cells) among CD200^high cells. Bars indicate mean ± SEM (**p = 0.005 by two-tailed Mann-Whitney test, n = 6 smears from two biopsies). c Representative images of cryosection of normal breast stained by immunofluorescence for CD200 (green), K17 (red), and nuclei (blue) show co-localization of CD200 and K17 in the terminal duct (TD) rather than alveoli (Alv) of a TDLU (encircled, scale bar, 100 μm).