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6 protocols using sodium chromate

1

Cell Culture Reagent Acquisition Protocol

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Sodium chromate, lead chromate, colcemid, and potassium chloride were purchased from Sigma-Aldrich (St. Louis, MO). Gurr’s buffer, trypsin/EDTA, sodium pyruvate, penicillin/streptomycin, and L-Glutamine were purchased from Invitrogen (Carlsbad, CA). Giemsa stain was purchased from Biomedical Specialties (Santa Monica, CA). Sodium dodecyl sulfate (SDS) was purchased from American Bioanalytical (Natick, MA). Crystal violet, methanol and acetone were purchased from J.T. Baker (Phillipsburg, NJ). Dulbecco’s minimal essential medium and Ham’s F-12 (DMEM/F-12) were purchased from Mediatech (Herndon, VA). Cosmic calf serum (CCS) was purchased from Hyclone (Logan, UT). Tissue culture dishes, flasks, and plasticware were purchased from Corning (Acton, MA)
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2

Immunofluorescent Analysis of DNA Damage

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Sodium chromate, Triton X-100, demecolcine and potassium chloride (KCl) were purchased from Sigma Chemical (St. Louis, MO). Giemsa stain was purchased from Ricca Chemical Company. (Arlington, TX). Crystal violet, sodium dodecyl sulfate (SDS), and acetic acid were purchased from J.T. Baker (Phillipsburg, NJ). Nitric acid and methanol were purchased from BDH Chemicals (Radnor, PA). Gurr’s buffer, Keratinocyte-SFM, and 4′,6-diamidino-2-phenylindole (DAPI) were purchased from Invitrogen Corporation (Grand Island, NY). Tissue culture dishes, flasks and plasticware were purchased from BD (Franklin Lakes, NJ). Urothelial cell medium, trypsin/EDTA and trypsin neutralizing solution were purchased from ScienCell Research Laboratories (Carlsbad, CA). Penicillin/streptomycin was purchased from Mediatech (Manassas, VA). Paraformaldehyde 4% was purchased from Alfa Aesar (Ward Hill, MA). Phospho-histone H2A.X (Ser139) antibody was purchased from Cell Signaling (Beverly, MA). AlexaFluor 488-conjugated goat anti-rabbit IgG secondary antibody was purchased from ThermoScientific (Rockford, IL).
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3

Synthesis of Metal-Containing Compounds

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FeS (−100 mesh, 99.9%) and elemental Zn [Zn(0), −100 mesh, >90%] were purchased from Sigma-Aldrich (Milwaukee, WI, USA) and Kanto Chemicals (Tokyo, Japan), respectively. DNT (97%), 2,4-diaminotoluene (DAT, 98%), DCP (99%), 2-chlorophenol (2CP, 99%), 4-chlorophenol (4CP, 99%), phenol (98%), sodium chromate (Na2Cr2O4, 98%), sodium selenate (Na2SeO4, >95%), sodium selenite (Na2SeO3, 99%), HEPES (N-[2-hydroxyethyl]piperazine-N′-[ethanesulfonic acid]), and N-methyl-2-pyrrolidone (NMP, 99.5%) were purchased from Sigma-Aldrich. Methanol (high performance liquid chromatography [HPLC]-grade) was purchased from Burdick & Jackson (SK Chemicals, Ulsan, Korea). CaCl2·2H2O (>74%), Na2CO3 (>99%), NaHCO3 (>99.5%), and KOH (>85%) were provided by OCI (Seoul, South Korea), Daejung Chemical (Gyeonggi, South Korea), Junsei Chemical (Tokyo, Japan), and DC Chemical (Seoul, South Korea), respectively. All chemicals were used as received.
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4

Seahorse Assay Reagents and Materials

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Sodium chromate, glutamine (for Seahorse tests), oligomycin, 2-deoxyglucose, rotenone, and antimycin A were from Sigma-Aldrich (St. Louis, MO, USA). Trypsin/EDTA, sodium pyruvate, penicillin/streptomycin, LHC media, and L-glutamine were from Thermo Fisher Scientific (Waltham, MA, USA). Dulbecco’s minimal essential medium and Ham’s F-12 medium (DMEM/F-12) were from Mediatech (Herndon, VA, USA). Cosmic calf serum was from Hyclone (Logan, UT, USA). Collagenase Type II was from STEMCELL Technologies (Vancouver, Canada) Tissue culture dishes, flasks, and plastic ware were from Falcon Labware (Becton-Dickinson, Franklin Lakes, NJ, USA). Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and XF base medium were from Agilent Technologies (Santa Clara, CA, USA).
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5

Cell Proliferation Assay Protocol

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RPMI was purchased from Mediatech (Manassas, VA). Penicillin/streptomycin, Gurr’s buffer, and trypsin/EDTA were purchased from Invitrogen Corporation (Grand Island, NY). Crystal violet, acetic acid, and methanol were purchased from J.T. Baker (Phillipsburg, NJ) and Fetal Bovine Serum (FBS) was purchased from Gibco Life Technologies (Grand Island, NY). Tissue culture dishes, flasks, and plasticware were purchased from BD (Franklin Lakes, NJ). Lead chromate, sodium chromate, potassium chloride (KCl) and demecolchicine were purchased from Sigma/Aldrich. Giemsa stain was purchased from Biomedical Specialties Inc. (Santa Monica, CA).
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6

Cell Staining and Microscopy Protocol

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All plasticware was purchased from BD Falcon. Dulbecco’s Phosphate-Buffered Saline (DPBS), RPMI media with Glutagro was purchased from Corning. Potassium chloride, demecolcine, lead chromate, and sodium chromate were purchased from Sigma/Aldrich. Crystal violet, methanol and acetic acid were purchased from JT Baker. Microscope slides were purchased from Thermo Scientific. Giemsa stain was manufactured by Rica Chemical Co. Fetal Bovine Serum (FBS), Gurr’s buffer, trypsin, penicillin-streptomycin and sodium pyruvate was purchased from GIBCO Invitrogen Corporation.
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