Figure 2
From: CD146 is required for VEGF-C-induced lymphatic sprouting during lymphangiogenesis
VEGF-C elicitation of different signaling cascades downstream of CD146 or VEGFR-3. (a,b) Dose and time effect of VEGF-C156S on HDLEC cells. Serum-starved cells were treated with VEGF-C156S for 20 min at different concentrations in (a) or with 100 ng/ml of VEGF-C156S at different time intervals in (b). Dimerization of CD146 and phosphorylation and expression of AKT, ERK and p38, elicited from VEGF-C156S, were analyzed by WB. (c) HDLEC cells, transfected with siRNA of control, CD146, VEGFR-3 or combination of CD146 and VEGFR-3, were treated with 100 ng/ml of VEGF-C156S for 20 min. (d–g) HDLEC cells, pretreated with SCH772984 or FHPI for 72 hr, were subjected to spheroid sprouting assay (d), proliferation assay (e), tube formation assay (f) and transwell migration assay (g). VEGF-C156S was applied at the indicated concentration. Data were expressed as means ± SEM from 3 independent experiments (n = 12 in each group of d–g). Significant difference was determined by Student’s t-test (*P < 0.05; **P < 0.01; ***P < 0.001; ns, no significant difference). Full-length blots are presented in Supplementary Fig. S4.
