Fig. 6.

Effects of siRNA-mediated Akt1 knockdown on endothelial cell migration and apoptosis. HUVECs were transfected with the indicated concentrations of siRNA targeted against Akt1 (siAkt1) or a random sequence (siSCR). 48 h after transfection, cells were harvested. a RT-PCR analysis showed a reduction of mRNA level of Akt1. Cycling conditions: 26 cycles for Akt1 and Akt2, 22 cycles for β-actin. A single graph from triplicate determinations showing identical results was shown. b Protein levels were analyzed by Western Blot with anti-Akt antibody or β-actin antibody. A single graph from three independent experiments was shown. HUVECs were transfected with 30 nM siRNA targeting Akt1 (siAkt1) or a nonsilencing siSCR for 24 h and then low serum (5%)- and growth factor-starved for 12 h. The siAkt1- or siSCR-transfected cells were exposed to A549 CM. c Quantification of the endothelial wound repair was assessed by wound-healing assay. d The number of viable cells was determined by XTT assay. e Apoptotic cells were assessed by FACS analysis. Results were shown for percentage of unstimulated control siRNA-transfected cells