Figure 1.

Effect of normal and mutant human hemoglobins on human pulmonary arterial endothelial cells (HPAEC). (A) HPAEC were exposed to ferrous forms of either HbA, HbS, or HbE at equimolar concentration (100 μM) for 12 h. Cell lysates were immunoblotted with primary antibodies against HO-1, H-ferritin, phospho-NF-κB-p65 and NF-κB-p65 proteins. Equal loading was confirmed by re-probing the blots against β-actin. Blots are representative of three independent experiments. (B) Levels of lipid hydroperoxides and (C) mitochondrial superoxide radical generation was measured in HPAEC following exposure to different Hb proteins as described in the methods section (N = 3). Values from different treatment groups were compared using Mann-Whitney U-test. ^*P < 0.05 vs. untreated control, ^#P < 0.001 vs. untreated control.