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. 2016 Sep 20;16:740. doi: 10.1186/s12885-016-2769-0

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© The Author(s). 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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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Fig. 4 — Colony formation of MBCDF’s subpopulations on soft agar. Anchorage-independent cell growth of B3, B6, C1, D5 (Group 1), B2, C9, and F3 (Group 2) subpopulations was evaluated by soft agar assay. A bottom layer of 0.5 % of agar in RPMI plus 10 % FBS was placed. The top layer contained 0.35 % agar in RPMI plus 10 % FBS and 2500 cell/plate. Colonies were analyzed after 15 days of culture. MBCDF and T47D were used as positive controls of colony formation. Agar without cells was used as negative control. a Representative picture of colony formation of each subpopulations are presented. b Cells were photographed and counted. The graph represents the mean ± SEM. Soft agar assay was performed three independent times in triplicate