Streptomycin 450 201 el

The rat thoracic aortic smooth muscle cell line A7r5 (#BNCC339542) was purchased from the cell bank of the Chinese Academy of Sciences, Shanghai, China. A7r5 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (11995065/11885092 from Gibco, CA, United States), which was supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin (450-201-EL from Wisent Inc., CA, United States), and 10% FBS (10099-141 from Gibco, CA, United States), after which the cells were seeded at 1.25 × 104 per well in 6-well culture plates and allowed to grow for 2 days in DMEM, then the cells were divided into four groups and treated with DMEM solution containing 5.5 mM glucose (Normal glucose), 33 mM glucose (High glucose), 33 mM glucose + ACHP (High glucose + ACHP) or 33 mM glucose + turine (High glucose + taurine), respectively (Wang K. et al., 2017 (link)), where ACHP was added at a final concentration of 10 μM (Ma et al., 2015 (link)) and taurine at a final concentration of 25 mM (Pan et al., 2010 (link)). In all experiments, cells were serum-starved for 12 h with DMEM containing 0.5% FBS and then subjected to the different treatments.

Rat aortic/thoracic vascular smooth muscle cells (A7r5) (CRL-1444; ATCC) were maintained in minimum essential Dulbecco’s modification Eagle’s medium (DMEM) (319–005; Wisent), which contained 10% FBS, 0.6 IU/ml penicillin (450-201-EL; WISENT), 600 μg/ml streptomycin (450-201-EL; WISENT) and non-essential amino acids (NEAA) (312–012; Wisent). Cells were grown on 10 cm petri dishes (353003; Corning), unless specified otherwise, at 37°C in a humidified atmosphere with 5% CO2. The cells were passed twice per week until confluence was attained and starved 24 hours prior to experiments because FBS contains AGE harboring CML adducts (Figure A in S1 File).