Figure 6.

Roles of endogenous Prox1 in HDLECs. (A) Expression of endogenous Prox1 (left; green) was examined by specific antibodies in HUVECs (top) and HDLECs (bottom), with counterstaining for nuclei with propidium iodide (PI, middle; red; and right, merge). Bars, 100 μm. (B–E) Effects of Prox1 knockdown on expression of GAPDH (B), Prox1 (C), VEGFR3 (D), and integrin α9 (E) were examined by quantitative real-time PCR analysis. A scrambled siRNA sequence (Ambion) was used as a negative control siRNA (siNTC). Each value represents the mean of triplicate determinations; bars, SD. (F) Effects of Prox1 knockdown on the chemotaxes of HDLECs toward VEGF-A (50 ng/ml) and VEGF-C (50 ng/ml). Cell migration was measured by Boyden chamber assay as described in Materials and Methods. HDLECs were transfected with scrambled siRNAs (siNTC) or Prox1 siRNAs and plated on Boyden chambers with indicated chemoattractants placed in the lower wells. Results are expressed as the ratio of number of migrated cells normalized to control (no attractant). Each value represents the mean of triplicate determinations; bars, SD.