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. 2020 May 14;127(4):466–482. doi: 10.1161/CIRCRESAHA.119.316463

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© 2020 The Authors.

Circulation Research is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 1. — GLUT1 (glucose transporter isoform 1) inhibition impairs endothelial cell (EC) glucose metabolism and proliferation but not migration. A and B, ¹⁴C-3-O-methylglucose (3MG) transport (Kruskall-Wallis test and Dunn multiple comparisons test; A) and glycolytic flux (1-way ANOVA and Tukey multiple comparisons test; B) in cells from a brain-derived EC line (bEND3) incubated with 20 nmol/L BAY-876 (N4-[1-[(4-cyanophenyl)methyl]-5-methyl-3-(trifluoromethyl)-1H-pyrazol-4-yl]-7-fluoro-2,4-quinolinedicarboxamide) and 20 µmol/L cytochalasin B (CYT B) vs control. C, Glycolytic flux in human brain microvascular ECs (HBMV ECs) and human retinal microvascular ECs (HRMV ECs) incubated with 20 nmol/L BAY-876 vs control (Student t test). D, Abundances of glycolytic intermediates in bEND3 cells incubated with 20 nmol/L BAY-876 vs control (Student t test or Mann-Whitney U test). E, AMP/ATP ratio in cells from a brain-derived EC line (bEND3) incubated with 20 nmol/L BAY-876 vs control (Student t test). F and G, Western blot of p-AMPK (phospho-AMP-activated protein kinase), AMPK (F) and p-S6K1 (phospho-S6 kinase 1), p-RPS6 (phospho-ribosomal protein S6), p53, and p21 (G) in bEND3 cells incubated with 20 nmol/L BAY-876 vs control (Student t test). H, Proliferation rate of bEND3 cells incubated with 20 nmol/L BAY-876 and 20 µmol/L CYT B vs control (Kruskall-Wallis test and Dunn multiple comparisons test). I, Representative pictures and quantifications of scratch wound closure in bEND3 cells incubated with 20 nmol/L BAY-876 and 20 µmol/L CYT B vs control in conditions with and without mitomycin C (mito C) pretreatment (2-way ANOVA and Tukey multiple comparisons test). J, Representative pictures and quantifications of sprouting human umbilical vein EC (HUVEC) spheroids incubated with 40 nmol/L BAY-876 vs control in conditions with and without mitomycin C pretreatment (2-way ANOVA and Tukey multiple comparisons test). Scale bar=500 µm (I) and 100 µm (J). 3PG indicates 3-phosphoglycerate; AMP, adenosine monophosphate; ATP, adenosine triphosphate; DHAP, dihydroxyacetone phosphate; F1,6BP, fructose 1,6-bisphosphate; F6P, fructose 6-phosphate; G6P, glucose 6-phosphate; GA3P, glyceraldehyde 3-phosphate; Lact, lactate; PEP, phosphoenolpyruvate; and Pyr, pyruvate. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001.