Dulbecco s modified eagle s medium dmem

HeLa human cervical carcinoma cells were incubated in Dulbecco’s modified Eagle’s medium (DMEM) (Wako Pure Chemical Industries, Osaka, Japan) supplemented with 10% fetal bovine serum (FBS) (Equitech-Bio, Kerrville, TX, USA). MT-2, Jurkat, and K562 human leukemic cells were incubated in RPMI 1640 (Wako Pure Chemical Industries) supplemented with 10% FBS. Peripheral blood mononuclear cells (PBMC) were incubated in RPMI 1640, and supplemented with 10% fetal bovine serum (FBS) (Biosera, Kansas City, MO, USA) [45 (link)]. All media were supplemented with 89 μg/mL streptomycin (Meiji Seika Pharma, Tokyo, Japan), and cells were incubated at 37 °C in a humidified atmosphere of 95% air and 5% CO₂. Growing cells were plated at 2 × 10⁴ cells/mL into 24-well microtiter tissue culture plates (Iwaki brand Asahi Glass Co., Chiba, Japan) and incubated for 48 h before the addition of the drugs (the optimal cell number for cytotoxicity assays was determined in preliminary experiments). Stock solutions (1 mM, 3 mM, 10 mM, and 30 mM) of the compounds and imatinib (Wako Pure Chemical Industries) were prepared in dimethyl sulfoxide (DMSO; Wako Pure Chemical Industries) and then added to fresh culture medium. The concentration of DMSO in the final culture medium was 1%.

[¹⁴C]Urate (55.0 mCi/mmol), [³H]guanine (10.7 Ci/mmol), [³H]hypoxanthine (27.0 Ci/mmol), [³H]thymine (65.0 Ci/mmol), and [³H]uracil (42.8 Ci/mmol) were obtained from Moravek Biochemicals (Brea, CA), [¹⁴C]inulin (1.9 mCi/g) was from American Radiolabeled Chemicals (St. Louis, MO), and [³H]polyethylene glycol 4000 (PEG 4000, 1.5 mCi/g) was from PerkinElmer Life Sciences (Boston, MA). Unlabeled urate, guanine, hypoxanthine, thymine, and uracil were obtained from Wako Pure Chemical Industries (Osaka, Japan), and Ko143 was from Sigma‐Aldrich (St. Louis, MO). Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Wako Pure Chemical Industries and Invitrogen (Carlsbad, CA), respectively. Mouse monoclonal antibodies for the tag peptides of DYKDDDDK (FLAG) and hemagglutinin (HA) were obtained from Wako Pure Chemical Industries (product numbers of 014‐21881 and 018‐22381, respectively, for the anti‐FLAG and anti‐HA antibodies), and a mouse monoclonal antibody for β‐actin (product number A5441) and horseradish peroxidase‐conjugated goat anti‐mouse IgG (product number A8924) were from Sigma‐Aldrich. All other reagents were of analytical grade and commercially obtained.

Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s F-10 medium were obtained from Fujifilm Wako Pure Chemical Corp. (Osaka, Japan). Penicillin/streptomycin and trypsin–EDTA were purchased from Thermo Fischer Scientific (Rochester, NY, USA). Cell culture equipment and rectangular 8-well plates were obtained from BD Biosciences (San Jose, CA, USA) and Thermo Fisher Scientific, respectively. SUMILON Cell-disk LF1 was from Sumitomo Bakelite Co. LTD (Tokyo, Japan). Insert chamber (Falcon 353,095–353,097) was purchased from Corning (Lowell, MA, USA) and an ultrasonic cutter (ZO-91, Echo Tech Co. Ltd, Japan) was used to make an incision (~ 3 mm) for conducting electricity at the sidewall located ~ 5 mm above the bottom (Fig. S1). The insert chamber was placed between the carbon electrodes, one of which directly faced the incision. Calf serum (CS) and fetal bovine serum (FBS) were obtained from BioWest (Nuaille, France). Matrigel was obtained from Corning (#354230, NY, USA). Unless otherwise noted, all chemicals were of the purest grade available from Sigma Chemical (St Louis, MO, USA) or Fujifilm Wako Pure Chemical Corp.