Fig 10.
Deletion of Cdc42 in ECs decreased the VEGFR2 protein level. (A) Primary ECs isolated from Cdc42flox/flox mouse hearts were infected by Ad-Cre or by Ad-LacZ as a control. Whole-cell lysates were prepared and were blotted with antibodies against VEGFR1, VEGFR2, VE-cadherin, PECAM-1, Cdc42, and vinculin, as indicated. (B) Quantitative analysis shows that the VEGFR2 level was significantly decreased in Cdc42-null ECs. (C through F) Sections of E9.5 control (C and D) and CEKO (E and F) embryos were stained by antibodies against VEGFR2 (green) and PECAM-1 (red). Blood vessels are indicated by arrows, and dashed lines indicate the boundary of the neural tube (NT). Bars, 25 μm. (G) The membrane surface proteins of primary mouse ECs were labeled with biotin and were pulled down using streptavidin-conjugated beads. The precipitated complex was then analyzed by Western blotting using anti-VEGFR2 antibodies. (H) The intensities of VEGFR2 75-kDa fragment bands from three independent experiments were quantitatively analyzed. *, P < 0.05.