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. 2021 May 10;12:2616. doi: 10.1038/s41467-021-22771-3

Fig. 1. VEGF treatment increases MAMs formation in endothelial cells.

Fig. 1

Human umbilical vein endothelial cells (HUVECs) were treated with vascular endothelial growth factor (VEGF; 30 ng/mL) for 0, 15, 60, and 180 min. a Representative transmission electron microscopy images of ER and mitochondrial morphology. Bar graph illustrating quantitation of ER length adjacent to mitochondria normalized by total ER length and by the mitochondrial perimeter. (n = 5 independent experiments). Scale bars, 500 nm. b Association between the endoplasmic reticulum (ER) and mitochondria (Mito) was analyzed by confocal microscopy. Representative confocal images are shown. Bar graph showing quantification of ER–mitochondria contacts using the Pearson’s coefficient. Scale bars, 10 µm (n = 5 independent experiments). c Western blot analysis of mitochondria-associated endoplasmic reticulum membrane (MAM)-related proteins in VEGF-treated HUVECs. Relative expression was quantified by densitometric analysis of the western blot assay (n = 5 independent experiments). d Western blot analysis of MAM-related proteins in endothelial MAM fractions prepared from VEGF-treated HUVECs. Relative expression was quantified by densitometric analysis of the western blot assay (n = 5 independent experiments). Statistical significance was assessed using two-tailed t-tests for two groups and one-way ANOVA with post hoc multiple comparisons test for comparing multiple groups. *p < 0.05 vs VEGF (0 min). All values are mean ± S.D.