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. 2017 Jun 7;7:2968. doi: 10.1038/s41598-017-02916-5

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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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ML290 effects on MMP-2 expression and TGF-β1-induced Smad2 and Smad3 phosphorylation in HCFs. ML290 (1 μM) promoted MMP-2 activity to an equivalent extent to H2 relaxin (0.1 μM) over 72 hours. In (A upper) a representative cropped zymograph (see Figure S4) of duplicate samples from two separate experiments; in (A lower) mean ± SE OD MMP-2, expressed as the ratio of that of in the untreated control group. **p < 0.01 versus untreated control group. In HCFs treated for 72 hours, ML290 (1 μM) inhibited TGF-β1-induced increases in p-Smad2 (B, n = 4) and increases in p-Smad3 (C, n = 4) to a level similar to that with H2 relaxin (0.1 μM). TGF-β1 (2 ng/ml) increased p-Smad2 (B) and p-Smad3 (C) compared to vehicle treated samples. Neither H2 relaxin (0.1 μM) nor ML290 (1 μM) had a significant effect on p-Smad2 or p-Smad3 in cells not treated with TGF-β1 (B,C). Data are mean ± SEM of 4 independent experiments performed in duplicate, **p < 0.01 and *p < 0.05.