HUVECs were purchased from the Institute of Basic Medicine, Chinese Academy of Medical Sciences (Beijing, China). LPS was acquired from Sigma-Aldrich (St. Louis, MO, USA). SRT1720 and EX-527 were purchased from MedChemExpress (Princeton, NJ, USA). Pue (purity ≥ 98%) was purchased from the Chinese Medicine Resource Center, Chinese Academy of Traditional Chinese Medicine (Beijing, China).
The cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco, Carlsbad, USA) containing 10% fetal bovine serum (Gibco) and 100 μg/mL penicillin-streptomycin (Gibco). Cells were passaged using trypsin-EDTA (Gibco). The cells were incubated at 37°C, 95% humidity, and 5% CO2. The medium was renewed every 2 days. HUVECs were used up to passage five [62 (link)]. The HUVECs were activated with 10 μg/mL LPS for 24 h [63 (link)] and pretreated with 10, 20, 50, 100, or 150 mg/L Pue for 24 h before LPS induction. As indicated, HUVECs were incubated with EX-527 (MedChemExpress) for 6 h to inhibit SIRT-1 activity or with SRT1720 (MedChemExpress) for 6 h to activate SIRT-1.
The cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco, Carlsbad, USA) containing 10% fetal bovine serum (Gibco) and 100 μg/mL penicillin-streptomycin (Gibco). Cells were passaged using trypsin-EDTA (Gibco). The cells were incubated at 37°C, 95% humidity, and 5% CO2. The medium was renewed every 2 days. HUVECs were used up to passage five [62 (link)]. The HUVECs were activated with 10 μg/mL LPS for 24 h [63 (link)] and pretreated with 10, 20, 50, 100, or 150 mg/L Pue for 24 h before LPS induction. As indicated, HUVECs were incubated with EX-527 (MedChemExpress) for 6 h to inhibit SIRT-1 activity or with SRT1720 (MedChemExpress) for 6 h to activate SIRT-1.
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