Fig. 3.

Cyanide-induced HIF-1α activation. (A) Cells were treated with 400 μM KCN (0.5–12 h), and cell lysates subjected to nuclear fractionation followed by Western blotting to determine HIF-1α levels. (B) Effects of antioxidants and a p38 inhibitor on HIF-1α protein expression. Cells were pretreated for 30 min with NAC (0.5 mM), CAT (0.2 mM), SB203580 (20 μM) or SB202474 (20 μM) prior to 400 μM KCN for 3 h. HIF-1α was determined by Western blotting. Densitometric data represent the mean±SEM of three separate determinations. (C) Cells expressing the HRE-luciferase reporter gene construct were exposed to KCN (400 μM) in the presence or absence of the compounds as indicated above. The reporter assay was conducted 3 h after cyanide addition. *Significantly different from control group; #significantly different from KCN alone group, P<0.05.