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. 2021 Mar 25;11:6838. doi: 10.1038/s41598-021-85331-1

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© The Author(s) 2021

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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Inhibition of ERK 1/2 activity prevents loss of tendon biomechanical properties in tendon explants cultured for 12 days. (A) Representative fluorescence microscopy images of tendon fascicles with nuclear staining of all cells (blue) and dead cells (EthD-1, violet). Scale bar 200 μm. (B) Quantification of cell viability in uncultured tendon explants (fresh tissue), methanol treated uncultured negative control tendons (devitalized tissue) and ex vivo cultured explants in reduced and standard culture treated without and with an ERK 1/2 inhibitor, N = 6. Devitalized tissue significantly differs from all other conditions. (C) Representative macroscopic images of tendon fascicles before treatment (uncultured tendon explants, day 0) and after 12 days of incubation in indicated culture conditions. Scale bar 10 mm. (D) Elastic moduli of isolated tendon fascicles (freshly isolated or cultured ex vivo in reduced conditions, standard conditions or treated with an ERK 1/2 inhibitor), N = 12 (N = 10 for standard culture, due to tissue rupture during the mounting procedure. Negative values of elastic modulus resulting from preliminary tissue failure were set to zero). The standard culture significantly differs from all other conditions. GraphPad Prism (version 8.4.3) was used to perform statistical analyses and generate the figures. Box plots (25th and 75th percentiles) with whiskers (5th and 95th percentiles), median (line) and mean (+). Bar plots represent mean values + SD. All statistical tests unless otherwise stated: Repeated measures ANOVA with Tukey's multiple comparisons test with *p < 0.05, ***p < 0.001, ****p < 0.0001.