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. 2017 Mar 2;8(20):32794–32806. doi: 10.18632/oncotarget.15825

Figure 1. Inhibition of phosphorylated-Smad1/5 and tube formation by metformin.

Figure 1

(A) Dose-dependent effect. HUVECs were treated with metformin (Met) at varying doses, followed by BMP9. (B) Time-dependent effect. HUVECs were treated with metformin for different time (hours), followed by BMP9. Cell extracts (20 μg) were blotted with antibodies, as indicated in A and B. (C) Inhibition of tube formation. HUVECs were seeded on Matrigel and treated with or without BMP9 (10 ng/mL) and/or metformin (10 mM) for 7 hours. The cells were stained with Calcein AM and photos taken under fluorescent microscopy. Representative images are shown. Scale bar: 100 μm. (D) Quantitative analysis. Tube formation was calculated by lengths using ImageJ program and plotted. Analysis was performed from duplicate experiments, from which 5 visual areas were taken. Significance was tested by one way ANOVA (average pixels ± SD, n=10). ** p<0.01.