In vitro test of in silico selected siRNAs molecules for targeting KDR. (A) Human umbilical vein endothelial cells (HUVECS) were seeded in 6 well dishes at a density of 50,000 cells/well and transfected with 20 nM siRNA and 1 µg/mL AtuFECT as indicated. Two days after transfection cells were lysed and KDR expression was evaluated by immunoblotting analysis with KDR antibodies. Several siRNAs reduced expression of KDR. Expression of the house keeping gene p110α was not affected the siRNA transfections and was used as loading control. (B) Dose-dependent inhibition of KDR protein expression by selected KDR siRNAs. HUVECS were transfected with 20, 10, 5 and 1 nM of the selected siRNAs and AtuFECT at a constant siRNA to lipid ratio of 20 nM siRNA and 1 µg/mL lipid. Strongest reduction of KDR expression was observed by treatment with siRNAKDR2. (C) siRNA sequences used in this study. Nucleotides with 2′Ome modification are depicted in bold letters, nucleotides with 2′-fluoro modification are underlined. Whole blots are shown in Supplementary Fig. S1.