Pcs 100 010

Normal Primary Human umbilical vein endothelial cells (ATCC® PCS-100-010™) were maintained in endothelial cell medium (ECM; Sciencell, USA) in humidified incubator with 5 % CO₂ and 95 % air at 37 °C until desired cell density was reached. Confluent cells cultured up to 3–7 passages were first starved in 0.5 % fetal bovine serum ECM for 24 h before exposure to AngII (Sigma; 10⁻⁷, 10⁻⁶, 10⁻⁵ mol/L) for another 24 h.
The SIRT3 siRNA used was synthesized by Shanghai GenePharma (Shanghai). Negative control siRNA, which does not target any known gene, was used as a negative control. Cells were transfected with the siRNAs by the Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions and examined by western blot analysis. Cells transfected with SIRT3 siRNA were then treatment with or without AngII (10⁻⁶ mol/L) for 24 h.

Human umbilical vein endothelial cells (HUVECs) were purchased from ATCC (PCS-100-010) and were cultured using Endothelial medium (ScienCell,1001) at 37 °C in a humidified incubator (95% air with 5% CO₂). Specially, the medium was made from 500 mL basal medium, 25 mL fetal bovine serum (FBS), 5 mL endothelial cell growth supplement (ECGS) and 5 mL penicillin/streptomycin solution. For cells experiment, HUVECs were cultivated on Ni SACs/N-C-based sensor which was kept in the incubator for 24 h to allow cells to adhesion. These loosely bounded HUVECs were washed with sterile PBS before electrochemically recording of NO release.