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. 2020 Nov 26;10:20598. doi: 10.1038/s41598-020-75833-9

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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 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 geometry on fibronectin micropatterns regulates SMYD3 distribution. (a) Myoblast C2C12 cells expressing SMYD3-HA-FLAG plated on fibronectin coated substrates. Scale bars: 20 µm. (b) Quantification of the nuclear:cytoplasmic (<I_Nuc>/<I_Cyto >) distribution ratio for SMYD3-HA-Flag for cells spread on non-patterned substrates, and nuclear:cytoplasmic ratio of SMYD3-HA-Flag as a function of cell aspect ratio for cells spread on non-patterned substrates. The red line represents a linear regression (n = 202). (c) Myoblast C2C12 cells expressing SMYD3-HA-FLAG were plated on fibronectin micropatterns with the same area (1200 μm²), but different aspect ratios; square (1:1, upper panel) or rectangle (1:5, lower panel). The micrographs show the fibronectin (FN) patterning, the nuclear DNA staining (DAPI), SMYD3-HA-Flag protein location and F-actin (SiR-actin). The SMYD3 nuclear:cytoplasmic distribution appears different between square and rectangle patterns. Scale bars: 20 µm. (d) Quantification of the nuclear:cytoplasmic (< I_Nuc > / < I_Cyto >) distribution ratio for SMYD3-HA-Flag over a range of pattern areas (1200–1800 μm²) and geometries: squares (1:1 aspect ratio, green), rectangles (1:5 aspect ratio, blue) and elongated rectangles (1:8 aspect ratio, grey).The median value of the ratio is 27–42% higher for cells plated on squares than on rectangle patterns. A Kruskal–Wallis test was performed between conditions with identical areas. (e) Quantification of the nuclear:cytoplasmic (<I_Nuc>/<I_Cyto>) distribution ratio for control eGFP transfected into C2C12 cells on square or rectangle patterns (1200 μm²). The eGFP nuclear:cytoplasmic distribution is slightly different (less than 3%) between square and rectangle patterns. A two-sample t-test was performed. n = number of individual cells measured. *p < 0.05, ***p < 0.001, n.s. not statistically significant.