Fig. 5. effect of vascular endothelial growth factor (VEGF) on the expression of early and late markers of apoptosis and/or necrosis by dendritic cell (DC) cultures. The cells were cultured as described in legend of Fig. 3 with the maturation step carried out for 24 h with prostaglandin E2 + tumour necrosis factor alpha in the presence (DC7V) of VEGF or in its absence (DC7). In additional cultures the maturation step was carried out for 48 h in similar conditions, identified respectively as DC8V and DC8. The cells were double-stained for Annexin V/propidium iodide (apoptosis and/or necrosis) or stained only for caspase-3. A: cells that stained for Annexin V only (early apoptosis); B: cells double stained for Annexin V and propidium iodide only (late apoptosis/necrosis); C: frequencies of cells staining for Annexin V/PI + Annexin indicating the total number of cells undergoing apoptosis (early and late) and necrosis; D: cells expressing caspase 3 active (late apoptosis).