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2 protocols using phospho jnk1 2

1

Potassium Dichromate Cytotoxicity Assay

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K2Cr2O7 (molecular weight, 294.18) was purchased from Duksan pure chemical Company Limited, (Ansan, South Korea). Penicillin-streptomycin solution, trypsin-EDTA solution, DMEM, and 1% antibiotic-antimycotic solution were obtained from Life Technologies GIBCO (Grand Island, NY, USA). Fetal bovine serum (FBS) and an in vitro toxicology assay kit were purchased from Sigma-Aldrich (St. Louis, MO). The antibodies used for immunoblotting were against phospho P53, phospho ERK1/2, total ERK1/2, total AKT1 (Cell Signaling Technology, Beverly, MA), phospho P38 (Santa Cruz Biotechnology Inc., Santa Cruz, CA), phospho AKT1, phospho JNK1/2, total P53, total JNK1/2, total P38, BAX, BCL2, CASP9, CASP3, PARP, beta-actin (Abcam, Cambridge, MA), and cytochrome c (ENZO Diagnostics Inc., Farmingdale, NY).
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2

Immunoblotting and Nitric Oxide Assay

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10-H2DA, 10-HDAA, SEA, and LPS (Escherichia coli 0111:B4) and alkaline phosphatase-conjugated secondary antibody (anti-rabbit IgG) were purchased from Sigma (St. Louis, USA). Primary antibodies against β-tubulin, phospho-ERK1/2, phospho-JNK1/2, phospho-c-Jun (pS63), phospho-IκBα (pS36), and phospho-p65 were purchased from Abcam (Cambridge, Massachusetts, USA). Primary antibody against phospho-p38 (Thr180/Tyr182) was purchased from Cell Signaling Technology (Danvers, MA, USA). Griess reagent, NaNO2, and other chemicals of analytical grade were purchased from Sangon Biotechnology, Co. Ltd. (Shanghai, China).
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