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. 2020 Aug 4;10:13077. doi: 10.1038/s41598-020-69805-2

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© The Author(s) 2020

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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Pancreatic β-cells respond to IL-17F signals via IL-17RA and -RC. (A) Total RNA was isolated from Min6, NIT-1, pancreatic islets and colon tissue and the expression of Il17ra-e measured by Taqman quantitative RT-PCR. (B) NIT-1 cells and primary NOD islets were stained with antibodies against IL-17RA, RC and RD and cell surface expression quantified using flow cytometry (black line = antibody stained, grey line = unstained control). (C) NIT-1 cells were stimulated with IL-17F or IL-17A for 2 h and Nfkbiz expression measured by Taqman quantitative RT-PCR (n = 4 for all groups), p < 0.05 (*), p < 0.001 (***) One-Way ANOVA with Bonferonni’s Multiple Comparison test. (D) Min6 cells were stimulated with IL-17F or IL-17A for 4 h and Nfkbiz expression measured by Taqman quantitative RT-PCR (n = 4 for all groups), p < 0.001 (***) One-Way ANOVA with Bonferonni’s Multiple Comparison test. (E) NOD islets were stimulated with IL-17F or IL-17A for 4 h and Nfkbiz expression measured by Taqman quantitative RT-PCR (n = 3 for all groups), p < 0.001 (***) One-Way ANOVA with Bonferonni’s Multiple Comparison test. (F) IL-17RA knockout (left panel) and RC knockout (right panel) NIT-1 cells were generated with CRISPR/Cas9 and stained with antibodies against IL-17RA (left panel) and RC (right panel) (black = parental control cell line, grey = receptor knockout cell line, light grey = unstained control). (E) Wildtype, IL-17RA KO and IL-17RC KO NIT-1 cells were stimulated with IL-17F or IL-17A for 2 h and Nfkbiz expression measured by Taqman quantitative RT-PCR (n = 2 for all groups), p < 0.05 (*), p < 0.01 (**) Two-Way ANOVA with Bonferonni’s Multiple Comparison test.