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. 2014 Dec 20;16:504. doi: 10.1186/s13058-014-0504-9

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© Verbeke et al.; licensee BioMed Central 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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.

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Transformed BPECs grown in pWIT medium form squamous tumors. Human mammary epithelial cells were infected with vectors expressing TERT, BMI1, MYC and ESR1 (3G-ER cells). A. Western blot showing expression of the transgenes in parental (NT) and transformed cells. B&C. Tumor formation two weeks after subcutaneous injection of 3G-ER cells into NSG mice (n = 2). B. Stereomicrograph showing tdTomato fluorescence from the tumor. C. Glandular regions express keratin 8/18; squamous regions express keratin 14. All of the cells are ERα + because one of the lentiviruses used to transform the cells expresses ESR1. D&E. At 3 weeks (D, n = 2) and 5 weeks (E, n = 5) after injection, the tumors are dominated by stratified cells undergoing terminal squamous differentiation leading to the formation of keratin pearls. Keratin pearls are marked by asterisks. Scale bars: B 1 mm; upper panels in C 500 μm; lower panels in C 200 μm; D&E 200 μm.