GSM9416764: HAEC cell, hypoxanthine day1 1; Homo sapiens; RNA-Seq
Human aortic endothelial cells (HAECs)and human aortic smooth muscle cells(HASMCs)wereobtained fom ATCC(Manassas, VA,USA).HAECSwere cultured inendothelial growth medium,andHASMCs were maintained in Dulbecco's ModifiedEagle's Medium (DMEM), both supplemented with 10%fetal bovine serum (HyClone, Logan, UT, USA),2 mMglutamine,100U/L penicillin,and 100g/mstreptomycin. All cells were cultured at 37 oC in ahumidified atmosphere containing 5% CO,. Cells wererandomly assigned to treatment with either PBS orangiotensin Il (Ang Il, 100 nM)for 24 hours, followedby exposure to DMSO,metabolites (hypoxanthine orpyroglutamic acid,20)，ormetabolites ininhibitors (BCX-combination with pharmacological1777,a hypoxanthine inhibitor,or Pro-GA,apyroglutamic acid inhibitor, 20 μM)for an additional 6hours. Metabolites and pharmacological inhibitors werefrom MedChemExpress (MCE)(Princeton, NI, USA).CCK-8:Cell viability was measured using the CellCounting Kit-8(CCK-8,AqLab,Beijing, China)following themanufacturer's instructions.BrieflyHASMCs were seeded in 96-well plates at a density of 5x 103 cells per well and allowed to adhere overnight.After the indicated treatments, 10 uL of CCK-8 solutionwas added to each well, and the plates were incubated at37 'C for 2 hours. Absorbance at 450 nm was recordedusing a microplate reader. Cell viability was expressed asa percentage relative to untreated control cells. RNA was harvested using Rneasy mini plus kit (Qiagen). 1.3 ug of total RNA was used forthe construction of sequencing libraries.