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. 2017 Jul 21;7:6113. doi: 10.1038/s41598-017-06124-z

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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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TGF-β1 enhanced the binding of IL-37 to HUVECs. (A) TGF-β1 facilitates the binding of IL-37 to HUVECs. HUVECs were incubated with 1 μM biot-IL-37 with/without indicated concentrations of TGF-β1. Bound biot-IL-37 was assessed by flow cytometry. Gray histograms indicated unlabeled cells. The mean fluorescence intensity (MFI) of bound IL-37 was quantified (n = 4). (B) ALK-1 mediates TGF-β-facilitated binding of IL-37 to HUVECs. Flow cytometry of HUVECs incubated with indicated cytokines or antibodies. Biot-IL-37 and TGF-β1 were used at 1 μM. Anti-ALK1 antibody (α-ALK1) and IgG were used at 10 μg/mL. (C) Control and ALK1 siRNA-transfected HUVECs were incubated with indicated proteins or antibodies (10 μg/mL). Biotinylated IL-37 (biot-IL-37) and recombinant TGF-β were used at 1 μM. Data are presented as mean ± SEM. **P < 0.01, ***P < 0.001.