Fig. 2.

AMPK activation was required for ciglitazone-induced eNOS expression. (A) In vitro cultured RMVECs were induced by ox-LDL (50 μg/ml) in a time-dependent manner, the phosphorylation level of AMPK was detected by Western blotting. (B) AMPK phosphorylation in RMVECs after induction with ox-LDL (50 μg/ml) for 24 hrs with or without ciglitazone (10 μmol/l) pre-treatment. (C) The effect of ciglitazone (10 μmol/l) on ox-LDL-induced eNOS expression with or without AMPK siRNA (and scramble siRNA as negative control). (D) The effect of ciglitazone (10 μmol/l) on ox-LDL-induced RMVECs apoptosis with or without AMPK siRNA (scramble siRNA as negative control) detected by TUNEL staining. (E) Relative quantification of nitric oxide production using nitrate reductase method in RMVECs under different treatments. (F) Representative images of cell ageing (up) and endothelial tube formation in Matrigel Matrix from ox-LDL-induced RMVECs with or without pre-treatment of ciglitazone and AMPK siRNA (scramble siRNA as negative control). *P < 0.05 versus Control group; ^#P < 0.05 versus ox-LDL treated group; ^§P < 0.05 versus ox-LDL and ciglitazone treated group (n = 5 separated experiments).