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. 2018 Jul 13;8:10672. doi: 10.1038/s41598-018-28770-7

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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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Time-lapse intra vital microscopy reveals specific EC migratory behaviour in correlation with blood flow direction and arterio-venous identity. (A) Live imaging of the remodelling vasculature of Claudin5-GFP:NG2DsRED mice at day 7 after suture implantation allows for simultaneous recording of ECs (green) and pericytes (red). The dashed box area is magnified in (B). (B,B’) Vessels imaged day 7 and 8 respectively, expose migrating (magenta and white arrows) and static ECs (asterisks), in relation to flow direction (dashed arrow). The vessel bifurcation (dashed line) serves as reference point. (C,D) Venules imaged day 7 and 8, following suture implantation, reveal migratory ECs (magenta arrows), static ECs (asterisks), ECs with non-identifiable direction (unclear, orange arrowheads) as well as blood flow direction (dashed arrows). (E) Percentage of total ECs in a migratory, static or unclear state, within lumenized vessels (n = 6 mice). (F) Fraction of ECs, within the group defined as migratory, that migrate along or against flow direction, in lumenized venules or arterioles. ***p < 0.001. (G) EC migration speed (all directions, pooled data from 9 mice) within lumenized venules (n = 69) and arterioles (n = 38). (H) EC migration speed within lumenized venules and arterioles in relation to flow direction. *p < 0.05.