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. 2020 Jul 29;10:12685. doi: 10.1038/s41598-020-69698-1

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© The Author(s) 2020

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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Overexpression of Abi1 in mammary epithelial cells induces the EMT and increases stem cell activity. (A) MCF10A human mammary epithelial cells were stably transfected with Abi1 (pCXN2-Abi1) or the vector alone (pCXN2). Live phase contrast micrographs show that overexpression of Abi1 converted MCF10A cells from an epithelial to a fibroblastic shape with prominent lamellipodia (arrowheads in the inset). (B) Control and Abi1-expressing MCF10A cells were subjected to immunoblot analysis. Actin served as a loading control. (C) RT-PCR showed that Abi1 overexpression increased the expression of EMT-inducing transcription factors and the ratio of CD44/CD24. (D) Ethidium bromide-stained gels were quantified by densitometry and plotted as a ratio to 18S. N = 3, *P < 0.05 vs cells transfected with pCXN2 only. (E) Live phase contrast micrographs show control and Abi1-overexpressing MCF10A cells cultured on Matrigel for 24 h and 8 days (insets). *Indicates cavity formation. (F) Abi1 overexpression increased mammosphere formation. N = 24, *P < 0.01. (G) Matrigel invasion assay showed increased cell migration to the bottom of the transwell after Abi1 overexpression. (H) Cells migrated to the bottom of the transwell were counted in 20 × field and plotted. N = 24, *P < 0.001.