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. 2017 Aug 15;7:8277. doi: 10.1038/s41598-017-08883-1

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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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Prematurity modulates a subset of SASP. (A) Analysis of factors secreted by CT- and PT-ECFC using antibody arrays. CT-conditioned media signals were used as the baseline. The heat maps key represents log2-fold changes of PT-media from the baseline. Signals higher and lower than the baseline are highlighted in yellows and blue respectively. Factors that differed significantly between the CT- and PT-media are represented by an asterisk (*). (B) An ELISA assay confirmed that IL6 levels were significantly increased in conditioned media from PT vs. CT cells (upper panel, N = 9 vs. 5). Correlation between the secreted IL6 levels and the senescence percentage of ECFC (lower panel). (C) The qNano profiles of the concentrations of unfractionated raw vesicles for CT and PT-media using NP150 and NP400 nanopores (N = 5 vs. 6). (D) Size distribution of endothelial vesicles from CT- and PT-media, as estimated by qNano. The figure depicts the diameter of the vesicles (in nm) versus the normalized concentration of vesicles (in particles/ml). 60-nm bin size. (E) Comparison of levels of Annexin-V⁺ EMP of CT- and PT-media (N = 11 vs. 16). Representative plots and quantification of EMP measurements by flow cytometry (size of 0.1–1 µm that stained positively for Annexin-V). (F) Correlation between Annexin-V⁺ EMP levels and the senescence percentage of ECFC (N = 20). The data for all the bar graphs are presented as the means ± SEM. (Statistical analyses: Mann Whitney, Unpaired t-test; *p < 0.05, **p < 0.01, ***p < 0.001).