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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 junction dynamics and VE-cadherin internalization. (A and B) Still images from live microscopy of HUVECs transduced with lentivirus coding for VE-cadherin-EGFP (A) or LifeAct-EGFP (B) for 10 min prior and after addition of 20 ng/ml of VEGFA; untreated and shikonin-treated cells are shown. Scale bar, 5 µm. (C) Number of inter-cellular gaps at junctions of VE-cadherin-transduced HUVEC along time. (D and E) Number of JAIL and of VE-cadherin plaques at the lateral junctions of LifeAct-EGFP and VE-cadherin-EGFP-transduced HUVECs, respectively; means ± SEM, n = 6 junctions for 2 independent experiments. (F) Immunofluorescence of VE-cadherin (red) and Hoechst (blue, nuclei) before or after acid wash in HUVECs transfected with control or PKM2 siRNA and treated with 20 ng/ml VEGFA for 15 minutes. Scale bar, 10 µm. (G) Internalized VE-cadherin positive area per number of cells in arbitrary units (A.U.); n = 3 independent experiments, **p < 0.01 by one-way ANOVA with Sidak post-test. See related Figure S6 and Movies S5–S8 and S11–S14.