Fig. 3.

Effect of O₂ concentration and metabolic inhibitors on AMPK activation in HUVECs. (A) HUVECs were incubated for 2 h at different O₂ concentrations (21%, 3%, 1.5%, and <0.5%) in a controlled hypoxic chamber, and AMPK activation was assessed by Western blotting. (B) AMPK activation by bioenergetic crisis under normoxic conditions. Phosphorylated AMPK (pAMPK) in HUVECs treated with different glycolytic and mitochondrial inhibitors (20 mM 2DG and 0.5 μM rotenone) was assessed by Western blotting. These are representative data from at least three independent experiments. The values in brackets are the estimations from densitometry analysis. Rot, rotenone; AMPK, loading control.