Fig. 4
DisBa-01 inhibits VEGFR2 and β3 phosphorylation after VEGF stimulation. HUVECs (5 × 105 cells/well) were seeded in 6-well plates and left to adhere at 37 °C, 5% CO2, overnight, followed by a period of 24 h of starvation in serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and DisBa-01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO2, followed by cell lysis. Twenty micrograms of protein from cell lysates were resolved by SDS-PAGE. Blots were probed with antibodies to a P-TY1054 + TY1059 VEGFR2, to b P-Ty773β3 and GAPDH, this last to normalize loading. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of phosphorylated VEGFR2/GAPDH and β3/GAPDH expression from three independent experiments. Values of *p < 0.05 were significantly different when compared to untreated (a), DisBa-01 (b) and VEGF (c) groups