Fig. 3

Effects of strain and VEGFR-2 activation or inhibition on Src activity. a HMECs and b HUVECs were treated with control, no treatment (NT) media, 25 ng/mL VEGF (V), 3 µM SU5416 (SU), or both VEGF and SU5416 (V + SU) for 72 h. Western blot analysis for total levels of Src was performed and quantified. *p < 0.05 versus NT, ^ p < 0.05 versus V, # p < 0.05 versus SU. c HUVECs in 3D fibrin gels with magnetic beads were treated with 25 ng/mL VEGF (V), DMSO as a vehicle control (Veh), or 3 µM SU5416 (SU) for 72 h. Some samples received no external magnetic stimulation (No Magnet) while other samples were cultured above a rotating magnetic field (Magnet) to generate matrix distortions around the cells. Src was analyzed via Western blot and normalized to GAPDH as a loading control. * p < 0.05. d HMECs were subjected to control (NT) conditions without strain and VEGF, VEGF only (25 ng/mL, V), strain only (ε), or strain plus VEGF (V + ε) for 5 or 15 min before Src analysis via Western blot. e HMECs were treated with combinations of strain, VEGF (25 ng/mL), SU5416 (3 µM), and a vehicle control for 5 or 15 min. Samples were analyzed through Western blot and stained for total Src. f, g Quantification of Western blots for total Src in HUVECs with same treatment groups outlined above. Western blots were normalized to β-actin. For d and f, + p < 0.05 versus 0 min NT, * p < 0.05 versus NT at same time point, ^ p < 0.05 versus V at same time point, # p < 0.05 versus ε at same time point. For e and g, + p < 0.05 versus SU5416 with same treatment, * p < 0.05 compared to NT at same time point, ^ p < 0.05 versus V at same time point, # p < 0.05 compared to ε at same time point. Data shown as average + SEM, n = 6 for all samples except for c, where n = 4 excluding SU + mag where n = 2. Groups b–g were compared with Kruskal–Wallis test, followed by post hoc Dunn’s tests. Group a was compared with ANOVA, followed by post hoc Tukey HSD tests