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5 protocols using deferasirox

1

Preparation of Iron Chelation Agents

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Deferoxamine (Novartis Pharma SAS) was dissolved in sterile distilled water and deferasirox (Selleckchem S1712), was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 300 and 50 mmol/L, respectively. Ironomycin was a kind gift of Raphaël Rodriguez team (11 (link)), this compound was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 10 mmol/L. Other reagents: erastin (Selleckchem S7242, 10 mmol/L in DMSO), ferrostatin-1 (Selleckchem S7243, 50 mmol/L in DMSO), Q-VD Oph (SelleckChem S7311, 10 mmol/L in DMSO, 30′ pretreatment), iron(III) chloride hexahydrate (31232 Sigma Aldrich, 0.1 mol/L in water, 4 hours after treatment), reduced gluthatione GSH (Sigma Aldrich G4251, 0.1 M in PBS) H2O2 (Sigma Aldrich 216763), mafosfamide (surrogate of cyclophosphamide, Santa Cruz ChemCruz SC-211761, 10 mmol/L in saline water), gemcitabine (Sellekchem S1149, 50 mmol/L in saline water), doxorubicin (Sellekchem S1208, 20 mmol/L in DMSO), CldU (Abcam ab213715, 20 mmol/L), IdU (Abcam ab142581, 2 mmol/L), 3-methyl adenine (Merck, #189490), β-mercaptoethanol (Sigma, M3148), staurosporine (Selleckchem, S1421), ketokonazole (Sigma, UC280), Avasimibe (from Sigma, PZ0190), ibrutinib (from Sellekchem S2680), entospletinib (from Selleckchem, S7523), venetoclax (from Sellekchem, S8048), AZD-5991 (from Selleckchem, S8643), and A1155463 (from TargetMol, T6748).
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2

Ferroptosis Induction in AML Cells

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MOLM-14, OCI-AML2, HL-60, SET2, MV4-11, K562, THP-1, UT7-EPO, SKM1, NB4 and KASUMI-1 AML cell lines were used. Patients provided written informed consent in accordance with the Declaration of Helsinki. Bone marrow (BM) samples were obtained from five patients with newly diagnosed AML (characteristics provided in the Supplementary Table S1). Cells were cultured in RPMI with glutamine (Gibco61870, Life Technologies® Saint Aubin, France) supplemented with 10% fetal bovine serum (FCS) and 4 mM glutamine. Ferric Citrate (FAC) was purchased from Sigma. DHA, Ferrostatin-1, Deferoxamin (DFO), Deferasirox (DFX), QVD-OPH, APR-246, VPS34-in1, erastin, FIN56, and RSL3 for the in vitro study were sourced from Selleckchem (Houston, TX). L -C-Propargylglycine (PPG), Chloroquine and doxycycline were obtained from Sigma–Aldrich (Saint-Louis, MO). FINO2 was purchased from Cayman Chemicals (Ann Arbor, MI).
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3

Deferasirox and Ciclopirox for Iron Assay

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Deferasirox (S1712) and ciclopirox (S2528) were purchased from Selleck, USA. FITC-labeled holo-transferrin was from Jackson ImmunoResearch, USA. Ferric citrate and lactate were from Sangon Biotech, China.
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4

Multimodal iron homeostasis analysis

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The following chemicals were used: Deferoxamine mesylate (BioVision, 1883-1000), ammonium ferric citrate (I72-500; Fisher Scientific), holo-transferrin human (Sigma, T0665), deferasirox (DFX; Selleckchem, S1712), ferrostatin-1 (Cayman, 17729), CQ (Sigma, C6628), FerroOrange (Sigma, SCT210), Mito-FerroGreen (Dojindo Molecular Technologies, M489), Vectastain ABC-HRP kit (Vector Labs, PK-4001), MOM Mouse Elite detection kit (Vector Labs, PK-2200), and DAB Substrate kit (Vector Labs, SK-4100).
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5

Procurement of Chemical Compounds

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Chemical compounds were purchased from Sigma (ciclopirox, gemcitabine, and hydroxyurea) and Selleck Chemical (deferasirox).
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