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Erastin

Manufactured by Targetmol
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Erastin is a small molecule compound that functions as an inhibitor of the system Xc- cystine/glutamate antiporter. It is used in research applications to study the role of this cellular pathway in biological processes.

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Lab products found in correlation

Am251, by Targetmol (1 mentions) Ml 210, by Targetmol (1 mentions) Fin56, by Targetmol (1 mentions) Ly294002, by Targetmol (1 mentions) Pd98059, by Targetmol (1 mentions) Erastin, by Merck Group (1 mentions) Liproxstatin 1, by Merck Group (1 mentions) D8418, by Merck Group (1 mentions) Liproxstatin 1, by MedChemExpress (1 mentions) Fetal bovine serum (fbs), by Procell (1 mentions) Dulbecco modified eagle medium (dmem), by Procell (1 mentions) Arpe 19 cells, by Procell (1 mentions) Malondialdehyde, by Solarbio (1 mentions) Bca kit, by Beyotime (1 mentions) Tublin, by Cell Signaling Technology (1 mentions) Micro reduced glutathione gsh assay kit, by Solarbio (1 mentions) Nissl stain solution, by Solarbio (1 mentions) Acsl4, by Santa Cruz Biotechnology (1 mentions) Micro malondialdehyde mda assay kit, by Solarbio (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)

6 protocols using erastin

1

Screening Small-Molecule Inhibitors in Cell Lines

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The small-molecule inhibitors were all purchased from Targetmol, Wellesley Hills, MA, USA including Lipid metabolism compounds library (L2510), CB1 antagonists (AM251, T1915; AM281, T2264; JD5037, T4453; Otenabant, T3530; TM3837, T8511; CB1-IN-1, T5996), ferroptosis inducers (erastin, T1765; RSL3, T3646; ML-210, T8375; FIN56, T4066; CIL56, T4309), PI3K inhibitor (LY294002, T2008) and MEK inhibitor (PD98059, T2623).
Li P., Lin Q., Sun S., Yang N., Xia Y., Cao S., Zhang W., Li Q., Guo H., Zhu M., Wang Y., Zheng Z, & Li S. (2022). Inhibition of cannabinoid receptor type 1 sensitizes triple-negative breast cancer cells to ferroptosis via regulating fatty acid metabolism. Cell Death & Disease, 13(9), 808.
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2

Erastin Cytotoxicity Assay in A549 and H1299 Cells

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Cell Counting Kit-8 (CCK-8) (TargetMol) assay was performed to measure cell viability according to the manufacturer’s instructions. Erastin (T-1765) was purchased from TargetMol. A549 and H1299 cells were seeded in 96-well plates at a density of 3,000 cells per well and incubated in a humidified cell incubator at 37°C, 5% CO2 for 24 h. The cells were then treated with Erastin of 0, 10, and 20 for 24, 48, and 72 h. The WST-8 reagent was then added and the 96-well plate was continued to incubate for another 2 h. The OD (optical density) values were measured at 450 nm with a microplate reader. The experiments in all groups were performed in triplicates and repeated 3 times.
Tian Q., Zhou Y., Zhu L., Gao H, & Yang J. (2021). Development and Validation of a Ferroptosis-Related Gene Signature for Overall Survival Prediction in Lung Adenocarcinoma. Frontiers in Cell and Developmental Biology, 9, 684259.
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3

Ferroptosis Inducers and Inhibitors

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We utilized erastin (T1765, TargetMol, Wellesley Hills, MA, USA) and RSL3 (T3646 TargetMol, Wellesley Hills, MA, USA) as ferroptosis inducers (FIs). Liproxstatin-1 (HY-12726, MedChemExpress, NJ, USA) and acetylcysteine (NAC; A9166, Sigma‒Aldrich, Merck, Burlington, MA, USA) were utilized to offset ferroptosis and oxidative stress. The reagents, including DFIQ, erastin, RSL3 and Liproxstatin-1, were dissolved in dimethyl sulfoxide (DMSO; D8418, Sigma‒Aldrich, Merck, Burlington, MA, USA) at concentrations of 10 mM, 20 mM, 10 mM and 10 mM, respectively.
Bow Y.D., Ko C.C., Chang W.T., Chou S.Y., Hung C.T., Huang J.L., Tseng C.H., Chen Y.L., Li R.N, & Chiu C.C. (2023). A novel quinoline derivative, DFIQ, sensitizes NSCLC cells to ferroptosis by promoting oxidative stress accompanied by autophagic dysfunction and mitochondrial damage. Cancer Cell International, 23, 171.
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4

ARPE-19 Cells Culture Protocol

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ARPE-19 cells were purchased from Procell Life Science & Technology Co. Ltd., China (#CL-0026; preservation institution: ATCC, CRL-2302, BCRC), cultured in 90% dulbecco’s modified eagle medium (DMEM, #PM150210; Procell) with 10% FBS (#164210-500; Procell) at 37°C and 5% CO2. Fer-1 (#T6500; Target Molecule Corp., Boston, MA, USA), erastin (#T1765; TargetMol), AS-IV (#HY-N0431; MCE, USA), and EX 527 (#T6111; TargetMol) were used in the experiments.
Tang X., Li X., Zhang D, & Han W. (2022). Astragaloside-IV alleviates high glucose-induced ferroptosis in retinal pigment epithelial cells by disrupting the expression of miR-138-5p/Sirt1/Nrf2. Bioengineered, 13(4), 8240-8254.
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5

Ferroptosis Regulation in Nervous System

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The chemical and reagents used were Nissl stain solution (cresyl violet, G1430, Solarbio), malondialdehyde (MDA) assay kit; glutathione (GSH) assay kit, dimethyl sulfoxide (DMSO), BCA kits (Beyotime Biotechnology, Beijing), rabbit antibodies against antiglutathione peroxidase 4 (GPX4, ABclonal, USA, A13309), anticystine/glutamate antiporter (SLC7A11, ab175186, Abcom, USA), ACSL4 (sc-271800, Santa Cruz Biotechnology Inc., USA), Nrf2 (ABclonal, USA), GAPDH (1 : 1000, Cell Signaling Technologies, United States, #5174), tublin (1 : 1000, Cell Signaling Technologies, United States). Micro malondialdehyde (MDA) Assay Kit (BC0025, Solarbio Science & Technology Co., Ltd., Beijing.), and Micro Reduced Glutathione (GSH) Assay Kit (BC1175, Solarbio Science & Technology Co., Ltd., Beijing). Iron Assay Kit was obtained from Nanjing Jiancheng Co. (A039-2, China). Reactive Oxygen Species Assay Kit was obtained from Beyotime Biotechnology (S0033, Beijing). Fetal bovine serum (FBS) was purchased from GIBCO Inc. (Logan, Utah, USA). Erastin (T1765, TargetMol, USA) and RSL3 (HY-100218A, MedChemExpress, USA) were also used.
Hu W., Chen M., Wang W., Huang F., Tian X, & Xie L. (2022). Pomelo Peel Essential Oil Ameliorates Cerebral Ischemia-Reperfusion Injury through Regulating Redox Homeostasis in Rats and SH-SY5Y Cells. Oxidative Medicine and Cellular Longevity, 2022, 8279851.
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6

Proteomics of Ferroptosis Regulators

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The following commercial antibodies and reagents were used in this study: GPx4 (Abcam ab125066 and Santa Cruz sc-166570), SHARPIN (Proteintech 14626-1-AP), HOIL-1 (Santa Cruz sc-365523), CYLD (Santa Cruz sc-74435), OTULIN (Cell Signaling Technology 14127), β-actin (TransGen Biotech HC201), SLC7A11 (Cell Signaling Technology 98051), FSP1 (Proteintech 20886-1-AP), SELN (Santa Cruz sc-365824), SELT (Abcam ab176192), Flag (Cell Signaling Technology 2368), HA (SAB T501), Myc (Sigma-Aldrich m4493), M1-Ub (EMD Millipore MABS451), K63-Ub (EMD Millipore 05-1308), K48-Ub (EMD Millipore 05-1307), Ub (Dako Z0458), His (TransGen Biotech HT501), Anti-Flag affinity gel (Sigma-Aldrich A2220), Anti-Myc affinity gel (Biotool B23402), Anti-HA affinity gel (Biotool B23302), Ni-NTA His-Bind Resin (EMD Millipore 70666-4), RSL3 (TargetMol T3646), Erastin (TargetMol T1765), CHX (MCE HY-12320), E64D (MCE HY-100229), PS-341 (MCE HY-10227), Fer-1 (MCE HY-100579), and Na2SeO3 (Sigma-Aldrich 214485).
Dong K., Wei R., Jin T., Zhang M., Shen J., Xiang H., Shan B., Yuan J, & Li Y. (2022). HOIP modulates the stability of GPx4 by linear ubiquitination. Proceedings of the National Academy of Sciences of the United States of America, 119(44), e2214227119.
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