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. 2019 Jun 24;9:9089. doi: 10.1038/s41598-019-45626-w

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© The Author(s) 2019

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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Validation of hIL-36γ binding and functional assays for compound triage. (A) Binding between Alexa-488 labeled hIL-36γ and hIL-36R/hIL-1RAcP ECD heterodimer was measured by FRET ratio (left panel, n = 3). Inhibition of interaction between labeled hIL-36γ and heterodimer was assessed by displacement using either unlabeled hIL-36γ (middle panel, n = 3) or hIL-36Ra (right panel, n = 2). (B) hIL-36γ function was measured by its ability to induce the secretion of IL-8 in human keratinocyte derived cell line HaCaT (left panel, n = 3). Ability to inhibit hIL-36γ function was assessed using hIL-36Ra (right panel, n = 3). (C) hIL-1β function was measured by its ability to induce the secretion of IL-8 in HaCaT cells, n = 3. (D) Human-to-mouse cross-species activity of hIL-36γ was measured by its ability to induce the secretion of CXCL1 from mouse NIH 3T3 cells, n = 2. (E) Specificity of cross-species activity of hIL-36γ was tested by functional inhibition using mouse (left panel, n = 2) and human (right panel, n = 2) IL-36Ra. All data represents an average ± S.E.M.