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2 protocols using cell culture plates

1

Apoptosis Induction in Cell Culture

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GOSS (purity ≥90%), doxorubicin (purity ≥99%), and cell culture flasks were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Trypsin-EDTA solution and Alamar Blue® (a solution of resazurin fluorescence dye) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dimethyl sulfoxide (DMSO), penicillin/streptomycin and Dulbecco's phosphate-buffered saline (DPBS) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). Dulbecco's modified Eagle's medium (DMEM), heat-inactivated fetal bovine serum (FBS), and cell culture plates were purchased from VWR International (Radnor, PA, USA). An Annexin V-FITC Apoptosis Detection Kit Plus (cat. no. 68FT-Ann VP-S) was purchased from RayBiotech (Norcross, GA, USA). A DNA-free™ kit (cat. no. AM1907) was purchased from Life Technologies, Inc. (Thermo Fisher Scientific, Inc., Waltham, MA, USA). An iScript™ cDNA Synthesis kit (cat. no. 170-8890), SsoAdvanced™ Universal SYBR® Green Supermix and the Human Apoptosis PCR array (SAB Target List) H96 were purchased from Bio-Rad Laboratories (Hercules, CA, USA).
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2

Inflammatory Cytokine Assay Protocol

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The compound GOSS (purity ≥90%) was purchased from Santa Cruz Biotechnology, Inc. Trypsin-EDTA solution 0.25% and Alamar Blue® (a sterile buffered solution of resazurin fluorescence dye) were purchased from Sigma-Aldrich; Merck KGaA. Dimethyl sulfoxide (DMSO), penicillin/streptomycin, and Dulbecco's Phosphate Buffer Saline (DPBS) were obtained from the American Type Culture Collection. Dulbecco's Modified Eagle Medium (DMEM), heat-inactivated fetal bovine serum (FBS), and cell culture plates were purchased from VWR International (Radnor). TNF-α, Human Cytokine Antibody Array kit (cat. no. AAH-CYT-1000), Human ELISA kits for C-C Motif Ligand 2 [CCL2, also known as monocyte chemoattractant protein-1 (MCP-1), cat. no. ELH-MCP1] and Interleukin-8 (IL-8, also known as CXCL-8, cat. no. ELH-IL-8) were purchased from RayBiotech. TURBO DNA-free™ kit (cat. no. AM1907) was purchased from Life Technologies, Inc. TRIzol® reagent was purchased from Invitrogen; Thermo Fisher Scientific. An iScript™ cDNA Synthesis kit (cat. no. 170-8891), SsoAdvanced™ Universal SYBR® Green Supermix (cat. no. 1725271), Human PCR primers (CCL2, IKBKE, IL-8, STAT3, MAPK1, MAPK3, CCDC88A, PIK3CD, and GAPDH) were purchased from Bio-Rad Laboratories, Inc.
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