Fig. 2

Strain treatment alters VEGFR-2 expression and phosphorylation patterns in HMECs and HUVECs. a HMECs were grown on Flexcell plates and 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. Western blots were performed to analyze total VEGFR-2, b pY1054/Y1059, or c pY1214 levels relative to the 0 min, NT control groups. d HMECs were treated with combinations of strain, VEGF (25 ng/mL), SU5416 (3 µM), and a vehicle control then stained for total VEGFR-2, e pY1054/Y1059, and f pY1214. g–l Quantification of Western blots for total or phosphorylated VEGFR-2 in HUVECs with same treatment groups outlined above; however, pY1054/Y1059 was quantified for the 160 kDa cleavage product due to partial absence of the mature product in the blot for part k. Western blot quantifications were normalized to β-actin, with phosphorylation quantifications being double-normalized to total VEGFR-2. For a–c and g–i, + p < 0.05 versus 0 min NT, * p < 0.05 compared to NT at same time point, ^ p < 0.05 versus V at same time point, # p < 0.05 compared to e at same time point. For d–f and j–l, + 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 a, d, g, and j, n = 3 for all other samples. Groups a, d–f, h–j, and l were compared with Kruskal–Wallis test, followed by post hoc Dunn’s tests. Groups b, c, g, and k were compared with ANOVA, followed by post hoc Tukey HSD tests