Propidium iodide apoptosis assay kit

Maduramicin ammonium (molecular weight = 934.16, purity > 97%, by HPLC) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution (5 mg/ml), aliquoted and stored at −20°C. Dulbecco’s modified Eagle’s medium (DMEM) and 0.05% trypsin-EDTA were obtained from Mediatech (Manassas, VA, USA). Fetal bovine serum (FBS) was from Atlanta Biologicals (Lawrenceville, GA, USA). CellTiter 96^® AQ_ueous One Solution Cell Proliferation Assay Kit was from Promega (Madison, WI, USA). CF488A-Annexin V and Propidium Iodide (PI) Apoptosis Assay Kit was purchased from Biotium (Hayward, CA, USA). Enhanced chemiluminescence solution was from Perkin-Elmer Life Science (Boston, MA, USA). The following antibodies were used: cleaved PARP, cleaved caspase 3, cleaved caspase 8, cleaved caspase 9, LC3A/B (Cell Signaling, Beverly, MA, USA), FasL, Fas/CD95, TNFα, TNFR1, TRAIL, DR4, DR5, FLIP S/L, FADD, TRADD, RIP, AIF, GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), p62/SQSTM1 (Novus Biologicals, Littleton, CO, USA), goat anti-mouse IgG-horseradish peroxidase (HRP), goat anti-rabbit IgG-HRP (Pierce, Rockford, IL, USA), and goat anti-mouse IgG1-Alexa Fluor^® 594 (Life Technologies, Carlsbad, CA, USA).