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. 2018 Jul 24;8:11142. doi: 10.1038/s41598-018-29465-9

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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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Dose response effect of PEDF on apoptosis, migration, and adhesion of mouse dermal endothelial cells (MDMEC). (A) Representative flow cytometric analysis of MDMEC exposed to PEDF or CPT. MDMEC were treated with PEDF (100 ng/ml) or CPT (5uM/positive control) for 24 hours. Early apoptotic cells were detected by flow cytometry using an annexin V apoptosis detection Kit. (B) Fas and caspase 3 mRNA expression after PEDF treatment. n = 3 for control Fas & caspase 3, and PEDF caspase 3; n = 6 for PEDF Fas. (C) In vitro migration of MDMEC following scratch wound placement with/without PEDF treatment. Wound area was quantified in pixels using ImageJ. Mean ± SD; n = 5 for 0 h, n = 5, 12, 12, and 15 for PEDF 0, 1, 100, and 500 ng/ml, respectively. (D) Representative images of cell migration in scratch wounds. Scale bar = 100 µm. (E) Adhesion of MDMEC with and without PEDF, assessed by measuring the quantity of remaining adherent cells following a 4 h exposure to PEDF. Mean ± SD; n = 3. **p < 0.01, ***p < 0.001, and ****p < 0.0001, Student’s t-tests was used for statistical analysis.