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. 2020 Jul 7;10:11133. doi: 10.1038/s41598-020-67814-9

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© The Author(s) 2020

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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Kras* effector blockade abrogates Kras*-mediated growth in 3D. (a,b) Brightfield images of iKras*p53* cells plated in 3D given either DMSO or a MEK inhibitor for seven days. Quantification of cluster area, 4 days post treatment; at least 100 clusters per treatment group were analyzed. Bars represent cluster area mean ± SEM. ANOVA statistical analysis; * indicates p < 0.01 of the cluster area. (c) Quantification of cluster cross sections with a single layer of epithelial cells adjacent to the Matrigel as well as lumen that accounts for > 75% of the cluster area. Over 50 cluster cross sections per treatment group were analyzed. Bars represent single layer cluster prevalence per technical replicate, mean ± SEM. ANOVA statistical analysis; * indicates p < 0.01. (d,e) Quantification of cluster cross sections of human UM5 cell lines and 4,668 cell lines. (f,g) Hematoxylin/eosin stains of representative cluster cross sections of of iKras*p53* cells plated in 3D given either DMSO or a MEK inhibitor for seven days. (h,i) Immunohistochemical staining of phosphorylated ERK at Thr202 and Tyr204 (pERK); brown dye indicates positive staining. Scale bars 50 µm.