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. 2015 Jan 27;15:7. doi: 10.1186/s12861-015-0058-9

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© De Silva et al.; licensee BioMed Central. 2015

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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Pregnancy induces proliferation in both the HR+ and HR- cells. (A) Representative images of carmine-alum-stained whole mounts of mammary glands from virgin, 3-day and 7-day pregnant FVB/N mice. Scale bar, 1 mm. (B) Confocal immunofluorescence of mammary gland sections from virgin, 3-day and 7-day pregnant mice stained for the proliferation marker 5-ethynyl-2’-deoxyuridine (EdU, green), Estrogen Receptor (ER, red) and the luminal cell marker cytokeratin-8 (CK8, blue). Scale bar, 10 μm. The exposure time for the ER signal was increased in pregnant samples to allow robust identification of ER+ cells (see Additional file 3 for images with a fixed exposure time). (C) Quantification of the proportion of luminal cells that is proliferating, separated by cell type based on ER expression. ER+ and ER- luminal cells start proliferating at day 3 of pregnancy, by day 7 the majority of proliferating luminal cells are ER-. Bars represent individual animals.