American Society of Hematology

Figure 4.

$Figure 4. Contribution of NRP-1 and VEGFR-2 to endothelial cell attachment. (A) Endothelial cell attachment to plates coated with BSA, laminin, fibronectin, and gelatin after 7 hours' incubation. HUVECs were transfected with control, VEGFR-2, or NRP-1 siRNA 48 hours earlier. Representative images from inverted microscopy (Olympus 1X51 microscope with 20 ×/0.40 Ph1 lens; Olympus Optical) display the fraction of cells that attached. Images were obtained with a Religa 1300 digital camera (Qimaging) using IPLab acquisition software (Scanalytics). (B) HUVEC attachment to wells measured by absorbance at 595 nM after staining the attached cells with crystal-violet. HUVECs were transfected with control, VEGFR-2, or NRP-1 siRNA 48 hours earlier and allowed to attach to coated wells for 7 hours. Representative experiment of 3 performed. (C) F-actin expression and distribution in HUVEC transfected with control, VEGFR-2, and NRP-1 siRNA, and subsequently allowed to attach for 4 or 24 hours on gelatin-coated dishes. Transfection was carried out 48 hours prior to attachment. Representative confocal images from 1 of 3 experiments performed.$

Contribution of NRP-1 and VEGFR-2 to endothelial cell attachment. (A) Endothelial cell attachment to plates coated with BSA, laminin, fibronectin, and gelatin after 7 hours' incubation. HUVECs were transfected with control, VEGFR-2, or NRP-1 siRNA 48 hours earlier. Representative images from inverted microscopy (Olympus 1X51 microscope with 20 ×/0.40 Ph1 lens; Olympus Optical) display the fraction of cells that attached. Images were obtained with a Religa 1300 digital camera (Qimaging) using IPLab acquisition software (Scanalytics). (B) HUVEC attachment to wells measured by absorbance at 595 nM after staining the attached cells with crystal-violet. HUVECs were transfected with control, VEGFR-2, or NRP-1 siRNA 48 hours earlier and allowed to attach to coated wells for 7 hours. Representative experiment of 3 performed. (C) F-actin expression and distribution in HUVEC transfected with control, VEGFR-2, and NRP-1 siRNA, and subsequently allowed to attach for 4 or 24 hours on gelatin-coated dishes. Transfection was carried out 48 hours prior to attachment. Representative confocal images from 1 of 3 experiments performed.

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