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. 2019 Aug 1;9:11187. doi: 10.1038/s41598-019-47593-8

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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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Spatial regulation of mitochondrial, metabolic, and metastatic pathways in micropatterned tumor-stromal assays (µTSA) assessed by RNA sequencing. (A) Schematics depicting the steps to create a MCF-7/BMSC µTSA; (B) Representative micropattern of an MCF-7 island (green: Pan-Cytokeratin) surrounded by BMSCs (red: Vimentin). Blue: nuclei. Scale bar: 100 μm. Cancer cells from the center and edge of the µTSA (opaque annular ring with dotted outlines) were isolated by laser capture microdissection¹⁷, and their RNA was extracted and sequenced; (C) Gene sets found to be enriched at the interface vs. center by RNA sequencing and gene set enrichment analysis (glycolysis has negative NES indicating enrichment in the center). (See Methods for more details. FDR: False Discovery Rate; FDR < 0.25 was considered significant.) MCF-7 cells from >6 micropatterns/experiment were microdissected and combined before RNA isolation and sequencing; N = 2 independent experiments.