Cre-regulated depletion of Rac1.

<p><b>A.</b> Wild-type primary endothelial cells were infected with pSico-Rac1 lentivirus. High-efficiency transduction was achieved as indicated by uniform GFP expression in infected cells (left panel). Cells sorted for GFP positivity and transfected with a Cre-recombinase expression plasmid (pTURBO-Cre) showed a significant loss of GFP (right panel). Scale bar: 10 µm. <b>B.</b> One week after pTURBO-Cre transfection, levels of mRNA, detected by semi-quantitative RT-PCR (upper panels), and protein, detected by Western blotting (lower panels), showed successful Rac1- and Flk-1-depletion in pSico-Rac1 and pSico-Flk-1 infected cells, respectively. Αctin RT-PCR and Western blotting for Hsc-70 were carried out to ensure equal RNA and protein loading, respectively. Bar graphs represent densitometric readouts of mRNA of relative Actin or Hsc-70 protein levels, respectively. *P<0.01. N = 3 independent experiments.