4 2 68 hydroxyethyl 1 piperazineethanesulfonic acid hepes

DEAE Sepharose Fast Flow, Sephadex G-75, and Coomassie brilliant blue G-250 were purchased from Solabio Technology Co. Ltd. (Beijing, China). Trichloroacetic acid (TCA), ethylenediaminetetraacetic acid disodium salt (EDTA), β-mercaptoethanol (β-M), and ammoniummum sulfate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Human vascular endothelial cells of the type EA.hy926 (CRL-2922) were purchased from American Type Culture Collection (Manassas, VA, USA). Dulbecco’s modified Eagle’s medium (DMEM), nonessential amino acids (NEAA), penicillin-streptomycin (pen-strep), fetal bovine serum (FBS), 4-(2-68 hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and 0.25% (w/v) trypsin-0.53 mM EDTA were purchased from Gibco Invitrogen (Burlington, ON, Canada). Dithiothreitol (DTT) and Triton-X-100 were obtained from Sigma-Aldrich (St. Louis, MO, USA). Tumor necrosis factor-alpha (TNFα) was obtained from R&D Systems (Minneapolis, MN, USA). Dihydroethidium (DHE) was purchased from Biotium (Fremont, CA, USA).

Peptides, including VRP, LKY, VRY, and V-F (purity > 98%) were synthesized by Genscript Corp (Piscataway, NJ, USA). Triton-X-100, dithiothreitol (DTT), and Ang II were obtained from Sigma (St Louis, MO, USA). Losartan potassium and dihydroethidium (DHE) were purchased from Tocris (Oakville, ON, Canada) and Biotium (Fremont, CA, USA), respectively. TNFα and A779 were obtained from R&D Systems (Minneapolis, MN, USA). Nonessential amino acids (NEAAs), penicillin−streptomycin, 4-(2−68 hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 0.25% (w/v) trypsin-0.53 mM ethylenediaminetetraacetic acid (EDTA), Dulbecco’s modified Eagle’s medium (DMEM), and fetal bovine serum (FBS) were purchased from Gibco Invitrogen (Burlington, ON, Canada). ECs (EA.hy926, CRL-2922) and VSMCs (A7r5, CRL-1444) were provided by the American Type Culture Collection (Manassas, VA, USA).

Cooked patties of beef hamburger and Beyond Meat hamburger were bought from A&W (Edmonton, Alberta, Canada), and cooked patties of Impossible Meat burger were bought from Burger King (Edmonton, AB, Canada). ACE (from rabbit lung), hippuryl-His-Leu (HHL), pepsin (porcine gastric mucosa), pancreatin (porcine pancreas), 2,4,6-trinitrobenzenesulfonic acid (TNBS), cytochrome C, aprotinin, vitamin B12, (glycine)₃, dithiothreitol (DTT) and angiotensin II (Ang II) were obtained from Sigma (Oakville, ON, Canada). Vascular smooth muscle A7r5 cell line was purchased from ATCC (Manassas, VA, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), 4-(2-68 hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and non-essential amino acids (NEAA) were obtained from Gibco Invitrogen (Burlington, ON, Canada). Dihydroethidium (DHE) was purchased from Biotium (Fremont, CA, USA). Solvents used for UPLC were of chromatographic grade. Other chemicals applied were of analytical grade.