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. 2017 Dec 13;7:17485. doi: 10.1038/s41598-017-17730-2

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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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The SOCS box is not essential for ChSOCS1 inhibition of JAK/STAT signalling in DF-1. (A) Schematic representation of the architecture of SOCS1 protein, expression plasmids encoding wild-type (SOCS1 WT) and SOCS box deletion mutant (SOCS1 ΔBOX) and alignment of sequences (in boxes) of the KIR motif and the SOCS box domain of human, mouse and chicken SOCS1 proteins. (B) Luciferase reporter gene assay in DF-1 for chicken Mx1 and viperin promoters following transient transfection expression plasmids SOCS1 WT and SOCS1 ΔBOX, each with and without chIFN-α treatment. Two-way Anova with Bonferroni posthoc test were used to analyse the data. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (C) Immunoblot confirming expression of Flag-tag in Flag-tagged expression plasmids encoding for wild-type (WT) SOCS1 and SOCS box deletion mutant (ΔBOX). No protein was observed when DF-1 were transfected with SOCS1 KIR deletion mutant (ΔKIR) plasmid. Panel (C) shows cropped images of the immunoblots; full-length blots are presented in Supplementary Fig. S5.