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. 2017 Jan 31;16:19. doi: 10.1186/s12943-017-0592-0

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© The Author(s). 2017

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. 3 — Notch promotes CCR7-mediated stemness. a Level of cyclic AMP (cAMP) in PyMT-Ccr7 ^WT mammospheres with and without CCL19 stimulation and Notch inhibition, determined by an AlphaScreen cAMP assay. Cells were treated with CCL19, CCL21 and RO4029097 for 30 min before analysis. b PyMT-Ccr7 ^WT secondary mammospheres were tested for phosphorylation of ERK as a read-out for CCR7 activation with and without ligand stimulation and Notch inhibition. Cells were treated with CCL19 and RO4029097 for 15 min before lysis for Western analysis. c Secondary mammosphere-forming efficiency (number of spheres/cells seeded) of PyMT-Ccr7 ^WT primary mammary tumor cells cultured with and without chemokines CCL19 and CCL21, and RO4029097. a–c n = 6 mice/experiment