Atherogenesis May Involve the Prooxidant and Proinflammatory Effects of Ferryl Hemoglobin
Figure 3
FerrylHb induces HO-1 and ferritin in EC. Confluent HUVECs grown on 6-well plates were exposed to heme, Hb, metHb, and ferrylHb (100 μmol/L heme or as indicated in complete medium containing 15% of FBS). After 4 hours of incubation total RNA was isolated and HO-1 mRNA level was measured by quantitative RT-PCR (panels (a) and (d)). For protein expression, HUVECs were solubilized after 8 hours of treatment. HO-1 and ferritin H and L expression was detected by Western blot ((b) and (e)) or with ELISA (c). Immunoblots were reprobed with GAPDH and are representative of three independent experiments. Results are shown as mean ± S.D. () from one representative experiment of three.
