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. 2023 Aug 4;25:91. doi: 10.1186/s13058-023-01690-9

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© The Author(s) 2023

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 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. 4 — miR-203 exposure in vivo on PyMT mice alters the expression of stem-like and differentiation markers in mammary tumors and fully prevents lung metastasis. A Left panel, Representative images of IHC staining for Ki67 (to test proliferation), CD44 and NeuN (as stem-like cell markers) in control and miR-203-treated tumors, at the experimental endpoint and after exposure to Dox on alternating weeks from tumor detection by micro-CT, as indicated in Fig. 3A. Right panel, Violin plots showing the quantification of markers staining. B Left panel, Representative images of IHC staining for H3K27me3, prolactin and progesterone receptor (PGR), to test evidences of differentiation on control and miR-203-treated tumors as in (A). Right panel, Violin plots showing the quantification of markers staining. C Illustrative H&E staining of lung macro- and micro-metastasis, found in several control mice at the experimental human endpoint. Representative examples are shown, from the 9 metastasis cases identified throughout the three in vivo experiments (depicted in Figs. 1, 2, 3). As shown in the table, the overall incidence of metastasis was 31,03% in control mice versus 0% in miR-203-treated mice. The bottom right panel shows a representative micro-CT image, pointing to one evident macro-metastasis (yellow arrow) found in a control mouse. Scale bar, 500 µm. In violin plots, six different fields from three independent tumor samples were analyzed. ****p < 0.0001; ***p < 0.001; n.s. not statistically different (Student’s t test)