Fig. 6.

uPARAP downregulation impairs the Crk/JNK/paxillin pathway. a, b, d–f LECs were treated with siRNAs targeting uPARAP (siU1 and siU2) or a control siRNA (Ctr) and stimulated for the indicated time (minutes) with VEGF-C. a–f Western blot of total and phosphorylated (p) VEGFR-2 (Y1175), VEGFR-3 (Y1230/31) (a), ERK, AKT, JNK (b, c), Crk (d, e) and Paxillin (Y118, S178) (f). c PAECs expressing uPARAP were transfected with a plasmid carrying an intact (WT) or mutated (VEGFR-3_Y1063/F) VEGFR-3 cDNA. JNK phosphorylation was evaluated by Western blot after VEGF-C stimulation. d, e LECs were treated (+ZM) or not (-ZM) with VEGFR-2 inhibitor (ZM323881), or siRNA against VEGFR-2 (siR2). For quantification of Western blot band densities, values were calculated as the ratio of phosphorylated to total form and normalised to the values obtained with the control (Ctr) (n = 3, biological replicates). Statistical analyses were performed using a non-parametric Mann–Whitney test. *P < 0.05