Fig. 1
DisBa-01 effects on VEGF-induced HUVEC viability, invasion, migration and adhesion. a Cells were treated with DisBa-01 (1000 nM), VEGF (10 ng/mL) or both proteins in DMEM supplemented with 0.5% FBS followed by 24 h of incubation. Cell viability was measured by spectrophotometry at 540 nm after incubation with MTT. b HUVECs (2 × 105 cells/well) were treated with 1000 nM DisBa-01 and/or VEGF (10 ng/mL) on serum-free DMEM for 30 min at 4 °C. Cells were pipetted into the Boyden’s chamber and then it was inserted on well containing DMEM 10% FBS. The negative control comprised of serum-free DMEM on the wells. Invasion was allowed to occur for 18 h at 37 °C. Cell nuclei were stained with DAPI (0.7 ng/μl). Quantification of invasive cells was measured by automated cell counting. c-d For the migration assay, HUVECs (1 × 105 cells/well) were exposed to DisBa-01 (1, 10, 100 and 1000 nM), VEGF (10 ng/mL) or VEGF plus DisBa-01 (1000 nM) and immediately inserted into the Boyden’s chamber. The chambers were immersed in 10% FBS medium and allowed to migrate for 6 h at 37 °C. Control chambers were inserted in serum-free medium. Cell nuclei were stained with DAPI (0.7 ng/μl) and cell migration was measured by automated cell counting. e-f HUVECs (1 × 105 cells/well) were treated with DisBa-01 (1000 nM) and/or VEGF (10 ng/mL) and were immediately incubated (37 °C, 1 h) in fibronectin and vitronectin precoated-wells. Negative control was comprised of wells coated with 2% BSA. Cell nuclei were stained with DAPI (0.7 ng/μl) and quantification of adhesion cells was measured by automated cell counting. Results represent the average of three independent experiments in triplicate. Values of *p < 0.05 were significantly different when compared to untreated (a), treated with DisBa-01 (b), or with VEGF (c)