Diaminofluorophore 4 amino 5 methylamino 2 7 difluorofluorescein diacetate daf fm da

Dulbecco’s modified Eagle medium (DMEM), Ham’s nutrient mixtures medium (F-12 medium), trypsin-EDTA, penicillin-streptomycin (P/S), and fetal bovine serum (FBS) were purchased from Gibco-BRL (Grand Island, NY, USA). PIP ELISA kit was purchased from TaKaRa Bio Inc (Shiga, Japan). The 2,7-dichlorofluorescein diacetate (DCFH2-DA), 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 1,3-Bis (diphenylphosphino) propane (DPPP), diaminofluorophore 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM-DA), and the ELISA kits used for the analysis of human MMPs were purchased from Sigma (St. Louis, MO, USA). All other chemicals used in this study were of analytical grade.

Fetal bovine serum (FBS) and Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Gibco (Rockville, MD, USA). 3-(4,5)-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), LPS, Griess reagents, dimethyl sulfoxide (DMSO), 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA), acridine orange, and diamino fluorophore 4-amino-5-methylamino-2′7′-difluorofluorescein diacetate (DAF-FM DA) were obtained from Sigma Aldrich (St. Louis, MO, USA). Phosphate-buffered saline (PBS), bovine serum albumin (BSA), and penicillin-streptomycin (P/S) were purchased from Solarbio (Beijing, China). ELISA kits for TNF-α, IL-1β, and PGE₂ were from Dakewei (Shenzhen, China). Primary antibodies against iNOS, COX-2, p38 MARK, phosphorylated p38 MARK, JNK, phosphorylated JNK, phosphorylated IKKα/IKKβ, ERK, phosphorylated ERK, β-actin, and the secondary antibody were obtained from Cell Signaling Technology Inc. (Boston, MA, USA). All other chemicals and reagents used in these experiments were analytical grade.