Figure 3.

Hypoxic exposure mediates neuroglobin (Ngb) oxidation and ferroportin ubiquitination leading to decreased Ngb and ferroportin expression along with release of cytochrome c (Cyt c). Representative western blots of (A) Ngb, (B) ferroportin, (C) Cyt c, (D) caspase–3, (E) oxidized Ngb, (F) ubiquitinated Ngb, (G) oxidized ferroportin, and (H) ubiquitinated ferroportin. Glyceraldehyde phosphate dehydrogenase (GAPDH) was considered as loading control for western blots. Optical density of the protein bands was determined and percentage of protein expression was calculated assuming normoxic values without NAC treatment to be 100%. Samples from which the protein of interest was immune-precipitated before western blotting were loaded to gels as negative control. ^*Denotes P<0.01 when compared with 0 day control and # denotes P<0.01 when compared with corresponding NAC-untreated group. H, hypoxia; NAC, N-acetyl cysteine; NC, negative control.