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. 2017 Apr 28;7:1253. doi: 10.1038/s41598-017-01324-z

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© The Author(s) 2017

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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Impaired ability to sustain high external force in Nespr-1^ΔKASH and LMNA^ΔK32 cells. (A) Confocal images of WT, Nespr-1^ΔKASH and LMNA^ΔK32 myoblasts on fibronectin-coated rigid matrix (glass) and stained for F-actin (phalloidin, red) and non-muscle myosin IIA (NM-2A, green). Nuclei are stained with DAPI (blue). Scale bar: 10 µm. (B,C) Zoom-in of actin cytoskeleton in the perinuclear regions. Confocal images were taken at the apical (B) and basal (C) surface of the cell. (D,E) Supranuclear actin cable number on hard surface in WT, Nespr-1^ΔKASH and LMNA^ΔK32 myoblasts. Values expressed as absolute values (D) and as percent changes versus values obtained at 12 kPa (E). Values are means ± SEM. n ≥ 20 in WT, Nespr-1^ΔKASH and LMNA^ΔK32 myoblasts, ***p < 0.01 vs WT. Only significant difference is figured (F–H) Cell matrix adhesions on hard surface. (F) Confocal images of WT, Nespr-1^ΔKASH and LMNA^ΔK32 myoblasts on hard matrix (glass) and stained with antibody against vinculin (green). Nuclei are stained with DAPI (blue). Scale bar: 10 µm. (G,H) Histograms of vinculin area in WT, Nespr-1^ΔKASH and LMNA^ΔK32 myoblasts, expressed as absolute values (G) and as percent changes versus values obtained at 12 kPa (H). At least 50 cells of each type were analysed. *p < 0.05 and ***p < 0.001 compared with corresponding value obtained on 12 kPa substrate. Only significant difference is figured.