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Fig. 4 | Respiratory Research

Fig. 4

From: VEGF isoforms have differential effects on permeability of human pulmonary microvascular endothelial cells

Fig. 4

HPMEC were stimulated with 20 ng/ml of VEGF165a, VEGF165b or without any stimulation (Ctl/control). Representative western blots showing VEGF stimulation at 5, 10 and 60 min plus loading control (α-tubulin) with densitometry (n = 3). a & b Western blot of phosphorylated MEK1/2 (a) and phosphorylated p42/p44MAPK (b) in HPMEC. Stimulation with VEGF165a increased phosphorylation of MEK1/2 and p42/p44 at 5 and 10 min in contrast with VEGF165b which induced increased phosphorylation only at 10 min for both proteins. c & d Measurement of cells resistance by Endohm system. Dotted lines correspond to cell pre-incubated with pMEK inhibitor UO126. UO126 inhibited the effect of VEGF165a (c) and the effect of VEGF165b (d). e Graphs represent the mean of the scratched area in HPMEC treated with UO126 inhibitor at 24 h relative to control. VEGF165a but not VEGF165b stimulation significantly diminished scratched area (p = 0.004) and was inhibited by UO126 (p = 0.03). Densitometry and scratch assay analysed by Kruskal-Wallis test with post hoc Dunn’s analysis. TEER measurement analysed using two-way ANOVA and Bonferroni post-test. All data were plotted as mean ± SEM (n = 3-6)

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