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. 2018 Apr 20;8:6331. doi: 10.1038/s41598-018-24576-9

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© The Author(s) 2018

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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KLK12 modulates fibronectin matrix assembly by endothelial cells. The impact of KLK12 on FN matrix assembly by human ECs was analyzed after treatment with 10 nM KLK12 for 4 h or 24 h in basal medium containing 500 nM FN or FN-EDB. (A) Immunofluorescence analysis. After treatment, cells and matrix were fixed, FN fibrils were immunodetected (monoclonal anti-FN antibody IST-2, green channel), and DNA was stained using Hoechst reagent (blue channel). Scale bar = 50 µm. (B) Western blot. After treatment, the DOC-insoluble protein fraction and conditioned media were harvested and analyzed by Western blotting. Upper panel: Detection of FN fibrils in the DOC-insoluble protein fraction using the IST-2 anti-FN monoclonal antibody. Lower panel: Detection of soluble FN in conditioned media using the FN9-1 antibody directed against the amino-terminal domains of FN. The sizes of the protein markers (in kDa) are indicated on the left.