DU145 and PC3 prostate cancer cell lines were cultured in RPMI 1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS; Sigma) and 1% penicillin–streptomycin (Corning). HeLa and 293T cell lines were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% FBS (Sigma) and penicillin–streptomycin (Corning). For SILAC labeling, the DU145 cells were maintained in RPMI 1640 medium for SILAC (Thermo), which was supplemented with 10% dialyzed FBS (Gibco), 1% penicillin–streptomycin, 25 mg/500 ml proline, and 50 mg/500 ml l -arginine and l -lysine (light) or 13C615N4-l -arginine and 13C615N2-l -lysine (heavy). The cells were labeled, respectively, in the light and heavy media for >6 generations before doing a further treatment. For hypoxia treatment, the cells were cultured in an incubator subchamber supplied with 1% O2/5% CO2/95% N2 and monitored by an oxygen sensor (BioSperix). The main chemicals used in this study included puromycin (EMD Millipore), MG132 (APExBIO), dimethyloxalylglycine (DMOG; APExBIO) and deferoxamine (DFO; Cayman).
Deferoxamine dfo
Manufactured by Cayman Chemical
Sourced in United States
Deferoxamine (DFO) is a chelating agent used in laboratory settings. Its core function is to bind and sequester metal ions, such as iron, in order to facilitate their removal from biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Ferrostatin 1, by Cayman Chemical (4 mentions)
Z vad fmk zvad, by Santa Cruz Biotechnology (3 mentions)
Vitamin e, by Fujifilm (2 mentions)
N acetyl l cysteine nac, by Merck Group (2 mentions)
Dialyzed fbs, by Thermo Fisher Scientific (1 mentions)
Silac, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium for silac, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicillin streptomycin, by Corning (1 mentions)
Puromycin, by Merck Group (1 mentions)
Mg132, by Apexbio (1 mentions)
Malonate, by Fujifilm (1 mentions)
Vitamin c, by Fujifilm (1 mentions)
Rotenone, by Merck Group (1 mentions)
Hoechst 33342 dye, by Merck Group (1 mentions)
Idebenone, by Cayman Chemical (1 mentions)
Chloroquine, by Merck Group (1 mentions)
6 protocols using deferoxamine dfo
1
Cell Culture and Isotopic Labeling for Proteomics
2
Evaluating Colorectal and Gastric Cancer Cell Death
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The HT29 human colorectal carcinoma cell line was purchased from Dainihon Pharmacy Co. (Tokyo, Japan). TMK-1 human gastric carcinoma cells were a gift from Professor Wataru Yasui (Molecular Pathology, Hiroshima University, Hiroshima, Japan), and the CT26 murine colon carcinoma cell line was a gift from Professor I. J. Fidler (MD Anderson Cancer Center, Houston, TX, USA). Cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum at 37 °C in 5% CO2.
BBR (25 μM for 48 h unless otherwise specified, Tokyo Chemical Industry Co., Ltd., Tokyo, Japan), 5-fluorouracil (5-FU), N-acetyl-L-cysteine (NAC 1 mM), rotenone (0.5 μM) (Sigma-Aldrich Inc., St. Louis, MO, USA), malonate (0.5 mM), vitamin E (20 μM) and vitamin C (1000 μM) (Fujifilm-WAKO Chemicals, Osaka, Japan), Z-VAD-FMK (ZVAD, 20 μM) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), ferrostatin-1 (FRS, 2 μM), deferoxamine (DFO, 200 μM) (Cayman Chemicals, Ann Arbor, MI, USA), were purchased from the manufacturers listed.
Cell death was detected using Live-or-Dye™ Fixable Viability staining kit (Cosmobio, Tokyo, Japan). Dead cells were counted by observation of 500 cells. Apoptotic figures were detected by Hoechst 33342 dye (Sigma) apoptotic figures were counted by observation of 500 cells.
BBR (25 μM for 48 h unless otherwise specified, Tokyo Chemical Industry Co., Ltd., Tokyo, Japan), 5-fluorouracil (5-FU), N-acetyl-L-cysteine (NAC 1 mM), rotenone (0.5 μM) (Sigma-Aldrich Inc., St. Louis, MO, USA), malonate (0.5 mM), vitamin E (20 μM) and vitamin C (1000 μM) (Fujifilm-WAKO Chemicals, Osaka, Japan), Z-VAD-FMK (ZVAD, 20 μM) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), ferrostatin-1 (FRS, 2 μM), deferoxamine (DFO, 200 μM) (Cayman Chemicals, Ann Arbor, MI, USA), were purchased from the manufacturers listed.
Cell death was detected using Live-or-Dye™ Fixable Viability staining kit (Cosmobio, Tokyo, Japan). Dead cells were counted by observation of 500 cells. Apoptotic figures were detected by Hoechst 33342 dye (Sigma) apoptotic figures were counted by observation of 500 cells.
3
Chemical Compounds for Cell Death Study
4
Cell Death Inhibitor Assay in H9c2 Cells
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For the cell death inhibitor assay, H9c2 cells were treated with human recombinant HMGB1 (40 μg/mL, Biolegend, San Diego, CA, USA) for 48 h, with or without the addition of cell death inhibitors. The following inhibitors were utilized: N-acetyl-L-cysteine (NAC, 1 mM, Sigma, St. Louis, MO, USA), Z-VAD-FMK (ZVAD, 20 μM, Santa Cruz Biotechnology, Santa Cruz, CA, USA), ferrostatin-1 (FRS, 2 μM), deferoxamine (DFO, 200 μM, Cayman Chemicals, Ann Arbor, MI, USA), and 4-phenylbutyric acid (4PBA, 200 μM, WAKO). Additionally, chloroquine (100 µM, WAKO) was employed to inhibit autophagy.
5
Ferroptosis Inhibition in Cell Viability
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ALA (SBI Pharmaceuticals Co., Ltd., Tokyo, Japan), N-acetyl-L-cysteine (NAC 1 mM) (Sigma-Aldrich Inc., St. Louis, MO, USA), vitamin E (10 μM), dimethyl sulfoxide (DMSO) (WAKO Chemicals, Osaka, Japan), Z-VAD-FMK (ZVAD, 10 μM) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), ferrostatin-1 (FRS, 1 μM), deferoxamine (DFO, 100 μM) (Cayman Chemicals, Ann Arbor, MI, USA), ZnPPIX and CoPPIX (Sigma), and CORM2 (WAKO) were purchased from the manufacturers listed.
6
Cobalt and Iron Chemical Synthesis
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Cobalt (II) chloride (CoCl2) and ammonium iron (II) sulfate hexahydrate (Fe2+)(FUJIFILM Wako Pure Chemical, Osaka, Japan), N-acetyl-L-cysteine (NAC, Sigma–Aldrich (St. Louis, MO) and deferoxamine (DFO, Cayman Chemical, Ann Arbor, MI, USA) were purchased.
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