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. 2017 Sep 8;7:11009. doi: 10.1038/s41598-017-11188-y

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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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Enhanced further metabolism of 15-epi-LXA₄ and MaR1 in diseased tendon stromal cells. Cells were incubated with 10 nM of 15-epi-LXA₄, MaR1 or vehicle for 24 h at 37 °C then with IL-1β for 24 h at 37 °C. At the end of the incubations 2 volumes of ice-cold methanol were added and products were identified and quantified using LM profiling (see methods for details). (A) MS-MS spectrum employed for the identification of 15-oxo-LXA₄ (left panel); and 15-oxo-LXA₄ concentration in stromal cell incubations from healthy volunteers (HV) and patients with tendinopathy (TD; right panel) (B) MS-MS Spectrum employed for the identification of 14-oxo-MaR1 (left panel); and 14-oxo-MaR1 concentration in stromal cell incubations from HV and TD (right panel). Statistically significant differences were calculated using pairwise Mann-Whitney U tests. Results are presented as means ± SEM. *P ≤ 0.05, n = 7 donors for HV and 6 donors for TD. (C) 15-PGDH mRNA expression in stromal cells derived from healthy hamstring (n = 5) and diseased supraspinatus tendons (n = 5) after stimulation with IL-1β (10ngml^—1) for 24 h. Statistically significant differences were calculated using pairwise Mann-Whitney U tests. Gene expression is normalized to β-actin, bars show median values. (D) Representative immunofluorescence images of tendon stromal cells isolated from healthy hamstring donors (n = 3) and patients (n = 3) with supraspinatus tendinopathy showing staining for 15-PGDH (red) and nuclei (cyan) after stimulation with IL-1β (10ngml^—1) for 24 h. (E) 15-PGDH mRNA expression in healthy subscapularis (n = 4 donors) and diseased supraspinatus (n = 14 donors) shoulder tendon tissues. Gene expression is normalized to β-actin, bars show median values. (F) Representative immunofluorescence images of sections of diseased and healthy shoulder tendons stained for 15-PGDH (red). Cyan represents POPO-1 nuclear counterstain. Scale bars, 20μm.