Reactive oxygen species (ROS) are associated with multiple cellular functions such as cell proliferation, differentiation and apoptosis. Signal transduction by ROS, ‘redox signaling’ in angiogenesis is an important and emerging topic in cardiovascular sciences. ROS mediate vascular endothelial cell (EC) proliferation and migration, but the molecular mechanism underlying this phenomenon remains largely unclear. Recently we investigated the pro-angiogenic effect of deoxyribose-1-phosphate (dRP) released by stimulated platelets, which was shown to induce an increase in EC motility in vitro and a strong angiogenic response in vivo. In this study, we investigated the role of ROS generation in the angiogenic response mediated by dRP. ROS generation was investigated in human umbilical vein endothelial cells (HUVECs) by live cell confocal imaging, which showed that the addition of exogenous dRP decisively increased ROS generation. Similarly, the dRP derivatives deoxyribose-5-phospate (dR5P), glyceraldehyde-3-phosphate (G3P), and ribose-1-phosphate (R1P) also induced ROS accumulation in HUVECs, whereas deoxyribose (dR) did not. As expected, the redox stress marker heme oxygenase-1 (HO-1) was shown to be upregulated by dRP and its derivatives. The expression of critical pro-angiogenic molecules such as vascular endothelial growth factor (VEGF), interleukin 8 (IL-8), and stromal cell-derived factor 1α (SDF-1α) in HUVECs treated with dRP was tested by immunoblot. Interestingly, integrin β3 was shown to be upregulated by HUVEC treatment with dRP and its phorphorylated derivatives (i.e. except dR), as demonstrated by immunoblot experiments in the presence of the ROS scavenger N-acetyl-cysteine (NAC) and the antioxidant apocynin. In parallel, only the phosphorylated derivatives of dRP appeared to increase HUVEC cell motility in scratch-healing assay. The causative link between integrin β3 upregulation and increased HUVEC motility was proved using integrin-specific inhibitory antbodies. In summary, this study demonstrates a link between dRP-dependent increase in endothelial ROS generation, the upregulation of integrin β3 and the increase in endothelial cell motility and angiogenic activity. The paracrine regulation of endothelial cell motility by platelet-derived dRP is likely to play an important role in the stimulation of postnatal angiogenesis and tissue repair.