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. 2018 Nov 19;8:17010. doi: 10.1038/s41598-018-34718-8

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© The Author(s) 2018

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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Activation of crucial signaling pathways in osteoarthritis after hydrostatic pressure (HP) stimulation. (A) Representative immunoblots from the same gel were cropped to show specific bands for phosphorylated and total protein of Akt, ERK1/2, p38, ribosomal protein S6 as well as β-Catenin using GAPDH as housekeeping protein. Full-length blots are presented in Supplementary Figure S2. Pellets of unstimulated and HP-stimulated groups were harvested once every week for the last 21 days of the experiment and protein was isolated. Both (B) ERK1/2 and (E) Akt showed a statistically significant increase in activation in HP-stimulated pellets compared to static controls at all timepoints. (C) Protein expression of p38 did not significantly change over time. (D) Ribosomal protein S6, a downstream target of ERK1/2 and mTOR, did not exhibit an enhanced activation at any sampling timepoint. (F) Expression of β-Catenin increased towards the end of the stimulation period for stimulated pellets whilst it declined for unstimulated pellets. Mean + SEM; data from 2 individual donors, 6 replicates for each donor; *p < 0.05.