Fig. 2 Self-organization into artificial mature blood vessels.

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Fig. 2 Self-organization into artificial mature blood vessels. Self-organization into artificial mature blood vessels. (A and B) Confocal imaging of vesseloid immunostainings at day 1. Nuclei are gray (DAPI), CD31 is blue (endothelial marker), and αSMA or tubulin labeling is orange. Images correspond to a maximal intensity projection along the z axis, except for the half panels indicated as “optical section,” which depict a representative image at the equatorial plane. Scale bars, 50 μm. (C) Fluorescence intensity profiles of αSMA and CD31. The four different measurements were aligned together based on the intersection between both intensity distributions, and this value was taken as the reference in the plot. Negative values indicate distance toward the alginate wall, and positive values indicate distance in the direction of the lumen. (D) Anchoring of SMCs in ECM. Immunostaining of laminin, CD31, and αSMA was performed. Images correspond to a projection of the z axis of each signal. Nuclei are in gray (DAPI), laminin in green, CD31 endothelial marker in blue, and αSMA in orange. Zoom of an equatorial section of the vesseloid. Scale bars, 100 μm. (E) KI67 nuclear signal and histogram representation of proliferation at days 1 and 5. (F) Activated caspase-3 signal and histogram representation of apoptosis at days 1 and 5. Scale bars, 50 μm. AU, arbitrary units; ns, not significant. Photo Credit: Laetitia Andrique (INSERM U1029), Gaelle Recher (CNRS UMR 5298). L. Andrique et al. Sci Adv 2019;5:eaau6562 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).