Celltiter 96 aqueous one solution cell proliferation analysis kit

Maduramicin ammonium (molecular weight = 934.16, purity > 97%, by HPLC) was 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). Okadaic acid (OA) was from Sigma (Saint Louis, MO, USA). CellTiter 96® AQ_ueous one solution cell proliferation analysis kit was from Promega (Madison, WI, USA). Enhanced chemiluminescence solution was from Perkin-Elmer Life Science (Boston, MA, USA). The following antibodies were used: ERK2, JNK, phosphorylated-JNK (p-JNK) (Thr183/Tyr185), c-Jun, p-c-Jun (Ser63), p38, p-PP2A (Tyr307), demethylated-PP2A (De-PP2A), HA and GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-ERK1/2 (Thr202/Tyr204) and p-p38 (Thr180/Tyr182) (Cell signaling technology, Beverly, MA, USA), PP2Ac, PP2A-A, PP2A-B and methylated-PP2A (Me-PP2A) (Upstate biotechnology, Waltham, MA, USA), PP5 (BD transduction laboratories, San Jose, CA, USA), FLAG (Sigma, Saint Louis, MO, USA), goat anti-mouse IgG-horseradish peroxidase and goat anti-rabbit IgG-horseradish peroxidase (Pierce, Rockford, IL, USA).