Elisa kits for pge2

To determine the secretion level of various cytokines, culture supernatants were collected from the hypoxic- or normoxic-cultured cells. ELISA kits for PGE₂ (R&D Systems, Minneapolis, MN, USA), PGI₂ (Cusabio, Wuhan, China) and VEGF (RayBiotech, Norcross, GA, USA) were used according to the manufacturer’s protocols.

Dulbecco’s Modified Eagle Medium (DMEM) and fetal bovine serum were purchased from Gibco (Grand Island, NY, USA). RPMI 1640 medium came from Cytiva (Marlborough, MA, USA), and penicillin/streptomycin antibiotics came from Invitrogen (Carlsbad, CA, USA). EZ-Cytox reagent was obtained from DoGenBio (Seoul, South Korea). Griess reagent, dimethyl sulfoxide (DMSO), protocatechuic acid, and lipopolysaccharide (LPS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The ELISA kits for PGE₂, TNF-α, IL-6, IL-8, and IL-1β were purchased from R&D Systems, Inc. (Minneapolis, MN, USA). Isookanin was obtained from ChemFaces Biochemical Co. Ltd. (Wuhan, Hubei, China), and dissolved in DMSO. The luciferase assay system and lipofectamine™ 3000 reagent were purchased from Promega Co. (Madison, TN, USA) and Invitrogen (Waltham, MA, USA), respectively. Mitogen-activated protein kinases (MAPKs) antibodies (p-ERK1/2, ERK1/2, p-p38, p38, p-SAPK/JNK, SAPK/JNK) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA), while secondary antibodies were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).