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. 2022 Jan 17;12:791. doi: 10.1038/s41598-021-03134-w

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© The Author(s) 2022

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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Quantification of global and local changes in collagen scaffold. (a) Gel compaction recorded longitudinally from day1 to day7."M" indicates MDA-MB-231 monoculture, "MF" indicates initial seeding concentration between MDA-MB-231 to NHLF is 1:1; "M3F" indicates initial seeding concentration between MDA-MB-231 to NHLF is 1:3; "M5F" indicates the initial seeding concentration between MDA-MB-231 to NHLF is 1:5; "MD" indicates MDA-MB-231 monoculture with 20µM GM6001 treatment; "MFD" indicates an initial seeding concentration ratio of 1:1 between MDA-MB-231 to NHLF with 20µM GM6001 treatment; "M3FD" indicates an initial seeding concentration ratio of 1:3 between MDA-MB-231 to NHLF with 20µM GM6001 treatment; "M5FD" indicates an initial seeding concentration ratio of 1:5 between MDA-MB-231 to NHLF with 20µM GM6001 treatment. (b) Quantification of gel compaction. The ratio indicates an initial seeding concentration between MDA to NHLF in each condition. Data from at least triplicates are averaged, and in each repeat there are duplicate wells for each condition. Error bars in SEM. (c) and (d) Pore size distribution on day 0 (c) and day 1 (d). Around 5–12 images (around 1000×1000 pixels) from two replicates were used for the quantification. The existence of NHLF contributes to the erosion of collagen scaffold and lead to larger hole size overall. GM6001 inhibits ECM erosion and breakage, consistent with the global gel compaction trend. (e) Binarized collagen images before pore size measurement. Similar to control conditions, day1 MDA monoculture generally has more small pores visually than coculture conditions, consistent with quantification shown in (d).