5 58 6 68 tetraethylbenzimidazolcarbocyanine iodide jc 1

Human umbilical vein endothelial cells (HUVECs) were obtained from ATCC. HUVECs were cultured with M199 basal medium supplemented with low-serum growth supplement and penicillin (50 IU/ml)-streptomycin (50 μg/ml). trypsin-EDTA was used to passage cells. M199 and trypsin-EDTA were obtained from Gibco (Grand Island, NY, USA). Low-serum growth supplement was purchased from Cascade (Portland, OR, USA). Additionally, 5,58,6,68-tetraethylbenzimidazolcarbocyanine iodide (JC-1) were purchased from BioVision (Palo Alto, CA, USA). LY294002, dihydroethidium (DHE), Apocynin, 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), DCHC, AICAR, sodium nitroprusside (SNP), Ro-320432, penicillin and streptomycin were all purchased from Sigma (St. Louis, MO, USA). Anti-β-actin, anti-AMPK, anti-AMPK-α, anti-AKT, anti-phospho AKT, anti-LOX-1, anti-SIRT1, anti-eNOS were all obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). HRP-conjugated anti-rabbit secondary antibodies were purchased from Transduction Laboratories (CA, USA). (TUNEL) staining kit was obtained from Boehringer Mannheim (Mannheim, Germany) was purchased from Selleckchem. Fura-2 AM and the EnzChek caspase 3 assay kit were purchased from Molecular Probes (Eugene, OR). SOD activity assay kit was obtained from Calbiochem (San Diego, CA). eNOS kit was purchased from R&D Systems (Minneapolis, MN, USA).