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. 2017 Dec 11;7:17297. doi: 10.1038/s41598-017-17609-2

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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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Ectodomain protein structures of grouper (Epinephelus spp.) toll-like receptor (TLR) 21s and activation of these TLRs by different CpG-oligodeoxynucleotides (CpG-ODNs). (A) Computational modeling of the ectodomain protein structures of orange-spotted grouper (osg, E. coioides) and giant grouper (gg, E. lanceolatus) TLR21s. From left to right: predicted ectodomain structure of osgTLR21, ggTLR21, and superimposition of these two ectodomains. N: N-terminal end, C: C-terminal end of the ectodomain. (B) Sequences of CpG-ODNs used in this study. (C) Relative luciferase activities of human embryonic kidney (HEK) 293 cells co-transfected with a control vector and expression vector for different grouper TLR21s as indicated, along with a nuclear factor (NF)-κB controlled luciferase reporter gene, and treated with 3 µM CpG-ODN for 7 h. Data represent means ± SD (n = 3 independent experiments). **P < 0.01 compared with the control. (D) Immunoblot analysis of the expression of the grouper TLR21s using β-actin as a loading control.