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. 2020 Nov 30;10:20869. doi: 10.1038/s41598-020-77842-0

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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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Cell sheet contraction promotes cytoskeletal rearrangements and pro-chondrogenic signaling compared to 2D cell culture. Representative confocal images of full dish (top–down) (a) 2D cultures and (b) 3D contracted sheets before chondrogenic induction at day-0 with phalloidin/F-actin (green) staining for cytoskeleton and DAPI (blue) nuclear staining. Quantitative real-time PCR gene expression for (c) cytoskeletal structure marker β-actin, (d) cell–cell marker β-catenin, and chondrogenic ECM markers (d) BMP2 and (f) COMP. All gene expression normalized to GAPDH and compared to the 2D cell culture 0-day control sample. Error bars represent means ± SD (n = 4) (*p < 0.05, **p < 0.01). Microscopic photos acquired with NIS Elements AR Software (v.4.30.01, https://www.nikon.com/products/microscope-solutions/support/download/software/imgsfw/nis-ar_v43001du164.htm) (a,b).