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. 2018 Jan 8;8:110. doi: 10.1038/s41598-017-18382-y

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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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Characterization of recombinant human DCN. (a) Ponceau-Red staining of a nitrocellulose membrane containing an IMAC purification of suspension media with 1% dialyzed FBS (negative control, NC), an IMAC purification of DCN-containing cell culture media with originally 1% dialyzed FBS (DCN), and the HiMark Pre-stained protein standard (PS). (b) The same membrane after discoloring and specific immunodetection of DCN. (c) Specific immunodetection of untreated DCN, deglycosylated DCN (DCN deglyc., 4 h and 20 h) and the SeeBlue® Plus2 Pre-stained protein standard (PS). (d) Schematic of a Co-IP for the protein interaction between DCN and TGF-β1. (e,f) Specific immunodetection of human DCN (e) or TGF-β1 from human platelets (f) show the Co-IP eluate (Co-IP), an unspecific background control without the DCN antibody (NC), the positive control (PC; 250 ng DCN or 100 ng TGF-β1) and the SeeBlue® Plus2 Pre-stained protein standard (PS). All blots are cropped, indicated by the black boxes. Full-length blots are presented in Supplementary Figure 7.