Perlecan Domain V Induces VEGf Secretion in Brain Endothelial Cells through Integrin α5β1 and ERK-Dependent Signaling Pathways
Figure 4
DV induces HIF-1α and increases eukaryotic initiation factor 4E.
(A) HIF-1α immunoblot (upper) and OD quantification (lower) following DV exposure. Note the transient HIF-1α band at 5 minutes. N = 3, *P<0.05, **P<0.01 and ***P<0.001 in comparison between vehicle and DV treated cells. (B) Immunoblots (upper) of eukaryotic initiation factor 4E phosphorylation (peIF4E) and total eIF4E after treatment of BECs with combinations of DV, LY294002 and AktIV as labeled. Cells were treated with DV for 5 min. OD quantification (lower) of peIF4E as normalized to total eIF4E. N = 3, **P<0.01 versus untreated cells, ##P<0.01 within LY294002 treated-group (C) Immunoblots (upper) of peIF4E and eIF4E from BECs treated with combinations of DV and U0126 as labeled demonstrating that U0126 significantly abolished DV-induced peIF4E hyperphosphorylation as OD quantified (lower). Cells were treated with DV for 5 min. N = 3, **P<0.01 versus control, ##P<0.01 versus DV-treated group. (D) Immunoblots (upper) of phosphorylated c-jun (pc-jun) and total c-jun from BECs treated with DV +/− U0126 demonstrating that U0126 significantly inhibited DV-induced c-Jun phosphorylation as OD quantified (lower). Note the significant increase after 30 min following DV treatment. N = 3, **P<0.01 in comparison to untreated cells, ##P<0.01 between U0126-treated and untreated cells.