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. 2016 Sep 29;18:97. doi: 10.1186/s13058-016-0757-6

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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 — Knockdown of galectin-3 (Gal3) reduces canonical Wnt and AKT signaling in CSC-like spheres. a Canonical Wnt signaling activity was evaluated by SuperTOP/FOP (STF) assays and normalized to Renilla luciferase (RLU = relative light units). Baseline Wnt activity was reduced in Gal3^negative GI-LM2G spheres (grey column) compared to Gal3^positive GI-LM2C spheres (white column). b Flow cytometric analysis of the Wnt target gene LGR5 in GI-LM2C (red) and GI-LM2G (green) spheres. Canonical Wnt activity in (a) correlated with LGR5 expression. c TGFβ signaling was evaluated by SBE4-luc assays measured as normalized firefly to Renilla luciferase activity (RLU = relative light units). GI-LM2C (red column) and GI-LM2G (green column) revealed no significant difference in TGFβ activity. d Western blot analysis shows typical results for EMT markers (E-cadherin, vimentin), Gal3 as well as phospho-AKT/total-AKT in GI-LM2C and GI-LM2G spheres. P-AKT was absent in GI-LM2G. Experiments were carried out in triplicate (n = 3)