Cell culture and chemicals

Human adrenocortical cell line, H295R (ATCC#: CRL-2128), was purchased from American Type Culture Collection. Cells were cultured in Dulbecco’s Modified Eagle Medium: Nutrient Mixture F-12 medium (GIBCO, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (GIBCO), 1% antibiotic solution, and 0.1% insulin, transferrin and selenium premix (BD Biosciences). Human embryonic kidney cell line, HEK293, was purchased from The Cell Bank of Chinese Academy of Science. Cells were cultured in DMEM (Gibco) supplemented with 10% fetal bovine serum and 1% antibiotic antimycotic solution. TTA-P2 was purchased from Alomone Laboratories; Angiotensin II and nifedipine were purchased from Sigma-Aldrich (Munich, Germany).