DFO, SIH, and ferritin increase DAF expression on vascular ECs. (A) HUVECs were left untreated or treated for up to 24 hours with CoPPIX (50 μM), and DAF mRNA levels were quantified by real-time PCR. (B) HUVECs were treated with desferrioxamine (DFO; 100 μM), SIH (100 μM), vehicle (UT), or transfected with adenoviruses expressing either heavy-chain ferritin (FHC) or βgal (MOI 200), for up to 48 hours before analysis of DAF expression by flow cytometry using mAb 1H4. (C) HUVECs were treated for 48 hours with VEGF (25 ng/mL) or TNF-α (10 ng/mL) and IFN-γ (500 U/mL) (T/I), in the presence or absence of DFO or vehicle alone (UT), and DAF expression was quantified by flow cytometry. Flow cytometric data are expressed as percentage increase in RFI above that of ECs treated with plain EC culture medium alone. (D) HUVECs were left untreated or treated for 6 hours with hemin (0.2 μM) in the presence or absence of DFO or vehicle alone (UT). DAF mRNA levels were quantified by real-time PCR. (E) HUVECs were left untreated or treated for up to 24 hours with VEGF (25 ng/mL), and DAF (▨) and ferritin (▩) mRNA levels were quantified by real-time PCR. Data are mean plus or minus SEM (n = 2-4 experiments). *P < .05; **P < .01.