Figure 1

DUSP3 is highly expressed in endothelial cells and its dowregulation inhibits in vitro tubulogenesis. (A) Immunohistochemistry of DUSP3 and Von Willbrand Factor (vWF) on paraffin embedded 4 μm serial sections of human cervix biopsies. Section 1 was stained with anti-vWF antibody and section 2 with anti-DUSP3 antibody. Sections 3 and 4 were stained with the secondary antibodies used to reveal vWF and DUSP3 staining respectively. (B) HUVEC cells were transfected with non-targeting siRNA (siCTL) or with DUSP3 targeting siRNA (siDUSP3-1 and siDUSP3-2). Efficiency of DUSP3 downregulation was measured at protein level using western blot 72 h after cell transfection. (Bi) Equal amount of proteins were resolved by SDS-PAGE, and western blot was performed using anti-DUSP3 antibody or anti-GAPDH as loading control. (Bii) Quantification and statistical analysis of DUSP3 protein expression in siCTL, siDUSP3-1 and siDUSP3-2 transfection conditions represented as a ratio of DUSP3 on GAPDH. (Ci) Phase contrast microscopy of siCTL, siDUSP3-1 and siDUSP3-2 transfected HUVECs seeded on pre-solidified Matrigel for 16 hours. (Cii) Quantitative analysis of the experiment shown in (Ci) obtained by measuring the tube lengths (left panel) and number of intersections (right panel) from 10 fields. *, P < 0,05 and **, P < 0.01.