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. 2017 Jun 29;7:4394. doi: 10.1038/s41598-017-03054-8

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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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Functional characterisation of ANGPTL6 variants location and functional effect (A) Human ANGPTL6 protein and location of the genetic variants. Schematic of the human ANGPTL6 protein with the three recognisable domains: signal peptide (SP) in green, coiled-coil domain (CCD) in purple, and fibrinogen-like domain (FLD) in yellow. As all genetic variants identified are located predominantly in the FLD, the partial tertiary structure of the C-terminal protein portion (amino acids 135–470), based on previously solved fibrinogen structure (1lwu) is presented. The single variant seen in controls only is shown in blue, variants identified in cases are in purple, the S284X variant is starred. (B) Functional characterisation of ANGPTL6 variants. Cells were transiently transfected with constructs encoding wild-type (WT) or mutant ANGPTL6; levels of protein secretion into the media were measured by ELISA; means+/− standard deviation (SD) for experiments performed in triplicate are shown (results were confirmed by Western blotting; data not shown). Statistical significance was measured using unpaired T-test with Welch’s correction using the GraphPad Prism software. *p < 0.05; ***p < 0.001.