3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt lps

Dulbecco’s modified Eagle’s medium (DMEM), antibiotics (penicillin and streptomycin), and trypsin-ethylenediaminetetraacetic acid (EDTA) were purchased from Gibco BRL (Grand Island, NY, USA). Fetal bovine serum (FBS) was obtained from Biowest (Kansas City, MO, USA), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), LPS, 4-nitrophenyl n-acetyl-b-d-glucosaminide (p-NAG), and monoclonal anti-DNP-IgE were supplied by Sigma–Aldrich (St. Louis, MO, USA). DNP-BSA was procured from Invitrogen (Gaithersburg, MD, USA). Primary antibodies against p-p38, p38, p-JNK, JNK, p-ERK, ERK, p-Lyn, Lyn, p-Syk, Syk, p-PLCγ, PLCγ, and β-actin were obtained from Cell Signaling Technology (Danvers, MA, USA), and FcεRIγ, from LSBio (Seattle, WA, USA). SensiFAST SYBR No-ROX kit mix was purchased from Biolines (Seoul, Korea), and human filaggrin, AQP3, and HA ELISA kits were obtained from CUSABIO (Seoul, Korea). Rat basophilic leukemia (RBL-2H3, ATCC^® CRL-2256) and murine macrophage (RAW264.7, ATCC^® TIB-71) cells were procured from the American Type Culture Collection (ATCC), while human immortalized keratinocyte (HaCaT) cells were obtained from Prof. Lee of Chosun University in Korea.

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.