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. 2019 Oct 21;9:15022. doi: 10.1038/s41598-019-50866-x

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© The Author(s) 2019

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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PKM2 is required for endothelial-cell junction remodeling during sprouting angiogenesis in vitro and in vivo. (A) Scheme illustrating the patching algorithm classification of VE-cadherin active (unstable)/inactive (stable) junctions adapted from Bentley et al., 2014. (B) Immunofluorescence of VE-cadherin (green, top) in 3D spheroid sprouts. Pseudo-colored images according to the patching algorithm classification are shown below. Scale bar, 10 µm. (C and D) Percentage of VE-cadherin active and inactive junction patches (C) and their ratio (D) in 3D spheroid formed by siRNA-silenced HUVECs; means ± SEM, n = 3 independent experiments. **p < 0.01 by Mann-Whitney test. (E) Immunofluorescence of VE-cadherin (green, top) in P6 mouse whole-mount retinas 72 hours after intraocular siRNA injection. Pseudo-colored images according to the patching algorithm classification are shown below. Scale bar, 10 µm. (F and G) Percentage of VE-cadherin active and inactive junction patches (F) and their ratio (G) in P6 mouse retinas 72 hours after siRNA injection; means ± SEM, n = 4 mice, *p < 0.05 by Welch’s test.