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. 2019 Aug 30;9:12607. doi: 10.1038/s41598-019-48816-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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3D spheroids exhibit differential mRNA and protein expression of markers of epithelial-mesenchymal transition (EMT), cell-cell contact, angiogenesis, and invasion/migration. Normalized mRNA levels of (a) WNT5A, TGFB1, TGFB2, (b) ITGA2, ITGA5, ITGB1, VIM, OCLN, CDH1, TJP1, CLDN1, CLDN4, (c) HIF1A, VEGFA, VEGFC, MMP9, and TIMP2, as measured using RT-qPCR. Significant differences between groups determined by unpaired t-test; n = 4. Significant differences between means were indicated by **(p < 0.01) or ***(p < 0.001). Non-significant differences indicated by n.s. (d) Cropped blots of MMP9 and TIMP2 protein bands as detected by Western blot, and Amido Black staining of total protein. n = 4. Membrane was cut below 63 kDa and top half used for the MMP9 blot. Remaining membrane was cut above 25 kDa and lower half used for the TIMP2 blot. Whole blots with protein ladders found in Supplementary Fig. S1.