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. 2020 Oct 28;10:18519. doi: 10.1038/s41598-020-74575-y

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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A549 epithelial cells and WI-38 fibroblasts co-cultured motility for the vertical configuration with cells moving along the length of the rectangular strip with a width of 100 μm in our HSH system. (A) The co-culture was imaged with phase contrast at 0 h and 48 h after being released. (B) Immunofluorescent images were captured after 24 h as well. A second representative response of the A549 epithelial cells and WI-38 fibroblast response at (C) 0 and 48 h, and (D) with immunofluorescence. (E) A diagram indicating the location of zone A and zone B on the HSH substrate for (F) quantifying the motility of the A549 epithelial cells and WI-38 fibroblasts on the vertical HSH substrate. Green pseudo-coloring for was for Cytokeratin and A549 epithelial cells and blue pseudo-coloring was for DAPI and the nucleus. The differences between the two cell types were statistically significant (P value = 10^–24) for the cells located inside the 100 μm wide soft strip (group A).