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. 2014 Oct 30;10(11):2036–2052. doi: 10.4161/auto.34398

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© 2014 The Author(s). Published with license by Taylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License http://creativecommons.org/licenses/by-nc/3.0/, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 3. — Monoallelic Becn1 loss results in functionally enriched mammary stem and progenitor cell populations. (A–C) Freshly isolated Becn1^+/+ (blue) and Becn1^+/− (red) MECs were used, experiments were performed 3 independent times and outcomes are presented as means ± SDs. (A) Primary mammosphere assays performed using Pi⁻ LIN(CD31,CD45,LY76)⁻ CD24⁺ CD29^hi MECs plated at 20,000 cells per well (left) and in limiting dilution of 500 to 25 CD24⁺ CD29^hi MECs plated (right) reveal increased MaSC activity in Becn1^+/− MECs. (B) 3D colony-formation by CD24⁺ CD29^hi Becn1^+/− compared with Becn1^+/+ MECs plated at 20,000 cells per well shows increased MaSC activity. (C) Colony formation by CD24⁺ CD29^lo Becn1^+/− compared with Becn1^+/+ MECs plated at 1,000 cells per well reveals increased colony forming ability in 2D-matrigel conditions. (D) Increased repopulation frequency and mammary fat pad filling are seen following transplantation of 250 CD24⁺ CD29^hi Becn1^+/− MECs isolated from 5-wk-old mice. Becn1^+/+ (left) and Becn1^+/− (right), contralateral transplantations in wild-type recipient mouse. Circles represent fat pads and the black color represents the percentage of the mammary fat pad that is filled in. (E) Representative whole mount images of outgrowths from contralateral CD24⁺ CD29^hi Becn1^+/+ and Becn1^+/− MEC transplantation are shown and reveal increased ductal side branching in Becn1^+/− samples. *P < 0.05 by a 2-tailed Student t test. Scale bar: (E) 5 mm for whole mounts; 2 mm for whole mount enlargements.