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. 2017 Nov 7;7:14761. doi: 10.1038/s41598-017-15348-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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5-HT7R engagement promotes the acquisition of a pro-fibrotic gene signature in human macrophages and contributes to pathology-associated parameters in a mouse model of skin fibrosis. (A) GSEA on the “t statistic-ranked” list of genes obtained from the 5-HT-treated M-MØ versus M-MØ limma analysis, using the “TGFβ signaling” gene set. Black vertical lines indicate the position of each of the genes included within the “TGFβ signaling” Hallmark gene set. (B) Relative expression of TGFB1 mRNA in non-treated M-MØ (-) or M-MØ exposed to 5-HT (5HT, 6 h) in the absence or in the presence of the 5-HT7R antagonists SB269970 or SB258719. Results are expressed as the TGFB1 mRNA level relative to the TBP mRNA level in the same sample (n = 6; *p < 0.05). (C) Relative expression of TGFB1 mRNA in non-treated M-MØ (-) or M-MØ exposed to 5-HT (5HT, 6 h) in the absence or in the presence of the PKA inhibitor RP8. Results are expressed as the TGFB1 mRNA level relative to the TBP and HPRT1 mRNA level in the same sample (n = 6; *p < 0.05; **p < 0.01). (D) Production of TGFβ1 by non-treated M-MØ (-) or M-MØ treated with 5-HT (5HT) for 24 h. (n = 3; **p < 0.01). (E) Fibrosis was measured as the collagen Masson stained area (upper panel), immunohistochemistry analysis of activated fibroblasts (α-SMA⁺, middle panel) and number of F4/80⁺ cells per area (lower panel), in lesional skin from saline- or bleomycin-treated control and Htr7 ^−/− mice. Shown are the mean and SEM of three independent experiments with 10 mice per group. Statistical significance was evaluated using Mann-Whitney U-test, (*p < 0.05; **p < 0.01). Representative skin sections stained are shown (bar, 50 μM).