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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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IL-17F induced cell death is iNOS dependent in mouse islets. NOD islets were stimulated with IL-17F or IL-17A in the presence of (A) IFNγ or (B) TNFα for 4 h and Nos2 expression measured by Taqman quantitative RT-PCR (n = 5 for all groups), p < 0.05 (*), p < 0.01 (**) One-Way ANOVA with Bonferonni’s Multiple Comparison test. (C) Min6 cells, (D) NOD islets and (E) C57Bl/6 islets were stimulated with IL-17F or IL-17A in the presence of TNFα + IFNγ (Cyto) for 24 h and Nos2 expression measured by Taqman quantitative RT-PCR (n = 4 for Min6 cells and C57Bl/6 islets, n = 6 for NOD islets), p < 0.05 (*), p < 0.001 (***) One-Way ANOVA with Bonferonni’s Multiple Comparison test. (F) NOD islets were stimulated with IL-17F or IL-17A and TNFα + IFNγ (Cyto) +/− Jnk inhibitor (SP600125) at 50 μM for 24 h and Nos2 expression measured by Taqman quantitative RT-PCR (n = 2 for all groups), p < 0.001 (***) Two-Way ANOVA with Bonferonni’s Multiple Comparison test. (G) NOD islets were stimulated with IL-17F or IL-17A and TNFα + IFNγ (Cyto) ± p38 inhibitor (SB203580) at 20 μM for 24 h and Nos2 expression measured by Taqman quantitative RT-PCR (n = 2 for all groups), p < 0.001 (***) Two-Way ANOVA with Bonferonni’s Multiple Comparison test. (H) NOD islets were stimulated with IL-17F or IL-17A and TNFα + IFNγ (Cyto) ± NF-κB inhibitor (BAY11-7082) at 10 μM for 24 h and Nos2 expression measured by Taqman quantitative RT-PCR (n = 2 for all groups), p < 0.01 (**) Two-Way ANOVA with Bonferonni’s Multiple Comparison test. (I) NOD islets were stimulated with IL-17F or IL-17A in the presence of TNFα + IFNγ (Cyto) for 4 days ± NMMA (1 mM) and cell death quantified by DNA fragmentation (n = 2 for all groups), p < 0.01 (**) One-Way ANOVA with Bonferonni’s Multiple Comparison test.