Fig. 3
Effect of AA and A2P on cobalt chloride induced HIF-1α protein accumulation in metastatic melanoma. WM9 metastatic melanoma cells were treated for 24 h with the hypoxia mimetic cobalt chloride (100 μM) in the presence or absence of AA and A2P and nuclear extracts analyzed by western blot. a The addition of AA (100–500 μM) reveals that 100 μM AA is unable to reduce CoCl2 induced HIF-1α accumulation. Higher concentrations of AA (250 and 500 μM) are necessary to reduce induced levels of HIF-1α b, while cells treated with A2P (5.0-100 μM) show that A2P efficiently reduces CoCl2 induced accumulation of HIF-1α at concentrations as low as 5 μM. Protein expression was normalized to β-actin. All treatments were repeated a minimum of 2 additional times with similar results