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. 2017 Dec 6;7:17080. doi: 10.1038/s41598-017-17013-w

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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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Glucose toxicity inhibited angiogenic capacity of iECs. (A) Capillary-like tubular structures of MS1 on Matrigel after treatment with 5.6 mM glucose (control), 35 mM glucose (HG group), 35 mM glucose plus 10⁻⁸ M insulin (HG + I group), 35 mM glucose plus 0.5 mM L-arginine (HG + R group) and 35 mM glucose plus 100 μM β-mercaptoethanol (HG + βME group) for 24 h (upper panel). Tubes and nets were automatically selected and marked in blue using Wimasis Image Analysis System (lower panel). Blue, the tubular structure. Red, tubes. White, branching points. (B–H) Quantitative analysis of tube formation capacity among groups. (B,C) The covered area (%) and total loop numbers were significantly decreased in HG group. (D,E) Total nets and relative total tube length (normalized against control group) in HG group were significantly higher than those in other groups. (F) Total branching points of capillary-like tubular structures of MS1 from control, HG, HG + I, HG + R and HG + βME group. (G) Relative mean loop area (normalized against control group) of MS1 in different groups. (H) Relative mean loop perimeter (normalized against control group) of MS1 in different groups. *P < 0.05 compared with HG group, ^# P < 0.01 compared with HG group.