Figure 6

Vegf165 attenuated hyperoxia-induced apoptosis in non-endothelial cells. FLK1-eGFP E11.5 lungs were harvested and exposed for 2 d. to (A) 3% oxygen, (B) 50% oxygen + isotype control, (C) 50% oxygen + Vegf165 (100 ng/ml) and (D) 50% oxygen + anti-NRP1 (10 microgram/ml) + Vegf165 (100 ng/ml), followed by staining for TUNEL (red) and DAPI (blue). Endothelial cells fluoresce green (GFP). Arrows indicate TUNEL positive cells. (E) Quantitation of TUNEL positive cells/total cells x 100 and (F) TUNEL positive, GFP negative cells/total GFP negative cells × 100. n = 4 separate experiments with 2–3 lungs/condition/experiment. *p <0.05 vs. 50% and 50% + Anti-NRP1 + Vegf165. Bar = 20 microns. Photographs at 40× magnification. White asterisks point airway lumen.