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. 2017 Sep 7;7:10806. doi: 10.1038/s41598-017-10521-9

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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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Primary hHSC and LX-2 cells have a profibrogenic phenotype and generate cAMP in response to H2-RLX and ML290. (a) Abundant α-SMA staining in hHSCs and LX2 cells indicating an activated myofibroblastic (profibrogenic) phenotype of these cells when cultured on plastic (scale bar = 50 μm). Relative mRNA expression (Ct values) of ACTB, COL1A2, MMP1 and RXFP1 in LX-2 cells (b) and hHSCs (c); values are the mean ± SD of three independent experiments (LX-2 cells) or three individual patient samples (hHSCs). (d) Representative H2-RLX (closed circles) and ML290 (open circles) dose responses in LX-2 cells (pEC₅₀ 8.8 and 4.0 respectively) following 40 h of culture on plastic and a 60 min agonist stimulation. (e) Representative H2-RLX (closed circles) and ML290 (open circles) dose response in hHSCs (pEC₅₀ 9.3, 5.8 respectively) following 40 h of culture on plastic and a 60 min agonist stimulation. (f) Representative ML290 analogues dose responses in LX-2 cells following 40 h of culture on plastic and a 60 min agonist stimulation.