erastin (Bio-techne, 5449); SAS (MedchemExpress, HY-14655); RSL3 (Cayman, 19288); FIN56 (Cayman, 25180); FINO2 (MedchemExpress, HY-129457); PANKi (Cayman, 31002); BSO (Sigma, B2515); DEM (Sigma, D97703); etomoxir sodium salt (Selleckchem, S8244); lovastatin (Selleckchem, S2061); TOFA (Selleckchem, S6690); dorsomorphin (Selleckchem, S7306); alisertib (Selleckchem, S1133); verdinexor (Cayman, 26171); leptomycin B (Cayman, 10004976); tipifarnib (MedchemExpress, HY-10502); methotrexate (Selleckchem, S1210); pitstop2 (Sigma, SML1169); EML425 (Selleckchem, S2977); C646 (Selleckchem, S7152); brequinar (Selleckchem, S3565); elamipretide (Selleckchem, S9803)
Methotrexate
Manufactured by Selleck Chemicals
Sourced in United States
Methotrexate is a laboratory chemical used in the synthesis and analysis of various organic compounds. It is a white to light yellow crystalline powder that is soluble in water and certain organic solvents. Methotrexate is commonly used as a reagent in various chemical reactions and analytical procedures, but its specific applications should be determined by qualified professionals.
Automatically generated - may contain errors
Lab products found in correlation
Cisplatin, by Selleck Chemicals (2 mentions)
Doxorubicin, by Selleck Chemicals (2 mentions)
Hypoxanthine, by Merck Group (2 mentions)
Thymidine, by Merck Group (2 mentions)
Cytarabine, by Selleck Chemicals (2 mentions)
Vincristine, by Selleck Chemicals (2 mentions)
Ibrutinib, by Selleck Chemicals (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Lovastatin, by Selleck Chemicals (1 mentions)
Erastin, by Bio-Techne (1 mentions)
Verdinexor, by Cayman Chemical (1 mentions)
Tipifarnib, by MedChemExpress (1 mentions)
Brequinar, by Selleck Chemicals (1 mentions)
Alisertib, by Selleck Chemicals (1 mentions)
Fin56, by Cayman Chemical (1 mentions)
Leptomycin b, by Cayman Chemical (1 mentions)
Pitstop 2, by Merck Group (1 mentions)
Dorsomorphin, by Selleck Chemicals (1 mentions)
Anlotinib, by Selleck Chemicals (1 mentions)
Lipofectamine, by Thermo Fisher Scientific (1 mentions)
10 protocols using methotrexate
1
Ferroptosis Inducers and Inhibitors Screening
2
Survivin's miR-596-binding site regulation
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U2OS, a typical OSA cell line, was purchased from the cell resources center of the Chinese Academy of Medical Sciences (Beijing, China). Five patient-derived cell lines (PDCs) were separated from the clinical specimens obtained from five patients with OSA during surgery as part of normal medical care. The lentivirus particles of pri-miR-596 or Survivin with a mutation of miR-596 targeted sequences located in 3′-UTR were constructed and purchased (Vigene Corporation, Jinan City, Shandong Province, China). The wild-type sequence or sequence with mutated miR-596-binding site of Survivin’s 3ʹUTR (1921–2100) was obtained by chemical synthesis and cloned into pGL4.26 plasmids to construct luciferase reporters by Vigene Corporation. The luciferase containing wild-type sequence of Survivin’s 3ʹUTR with miR-596-binding site was named as Luc, whereas the luciferase containing sequence of Survivin’s 3ʹUTR with mutated miR-596-binding site was named as LucMut. The inhibitor of the miR-596 was purchased from Thermo Fisher Scientific (Waltham, MA, USA). The antitumor agents doxorubicin (Selleck, TX, USA), cisplatin (Selleck), methotrexate (Selleck), or anlotinib (Selleck) were dissolved in DMSO and conserved in an −80°C condition.
3
Methotrexate and Lipofectamine Transfection
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Methotrexate (catalog: S1210) was obtained from Selleck Chemicals (Houston, TX, USA). The transfection reagent Lipofectamine (catalog: LMRNA001) was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
4
SHIN1-Mediated Metabolic Modulation
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The small molecules methotrexate (Selleckchem, S1210), hypoxanthine (Sigma, H9636), thymidine (Sigma, T1895), and sodium formate (Fisher Scientific, S648-500) were used. The dual SHMT1/2 inhibitor SHIN1 (Tocris, 6998) was used at the concentration of 10 µM. In total, 1 mM of sodium formate was used to demonstrate on-target effects of SHIN1. Glutamylcysteine synthetase inhibitor l -buthionine-sulfoximine (BSO; Sigma, 83730-53-4) was used at the concentration of 10 µM. Mitochondrial complex I inhibitor Piericidin A (Cayman, 15379) was used at the concentration of 0.1 µM. Brequinar (Cayman, 24445) was used at the concentration of 1 µm and 10 µM. Glucose-free media containing galactose was prepared by supplementing 25 mM galactose (Sigma, G5388) into glucose-free DMEM (#11966025, ThermoFisher) with 10% dialyzed FBS (#26400044, ThermoFisher). Cells were always treated with small molecules or special media 2 h prior to infection. Samples were harvested at 48 hpi.
5
Cytotoxicity Assay with Chemotherapeutics
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Raltitrexed, pemetrexed, and methotrexate were purchased from Selleck. 5FU, puromycin, choloroquine, dTMP and polybrene were obtained from Sigma (Saint Louis, MO). 3-(4,5-Dimethylthiazol-2-yl)−2,5-diphenyl tetrazolium bromide (MTT) was purchased from Amersco (Cat. 0793-1G, Solon, OH).
6
Drug Preparation and Characterization
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Dasatinib (S1021), Ibrutinib (S2680), MK-2206 (S1078), Cytarabine (Ara-C, S1648), Methotrexate (MTX, S1210), Gemcitabine (GEM, S1714), Topotecan (TPT, S1231), Doxorubicin (DOX, S1208), Vincristine (VCR, S1241), Cisplatin (CDDP, S1166), Oxaliplatin (OXA, S1224), Decitabine (DAC, S1200) were purchased from Selleck, USA.
All chemicals but CDDP and OXA were dissolved in dimethyl sulfoxide (DMSO, V900090, Merck, USA) to concentrations of 10mM and aliquoted and stored at −20°C. CDDP and OXA were dissolved in Dimethylformamide (DMF, D4551, Merck, USA) to concentrations of 10mM and aliquoted and stored at −80°C. Working concentrations for all chemicals were determined by lethal dose tests.
All chemicals but CDDP and OXA were dissolved in dimethyl sulfoxide (DMSO, V900090, Merck, USA) to concentrations of 10mM and aliquoted and stored at −20°C. CDDP and OXA were dissolved in Dimethylformamide (DMF, D4551, Merck, USA) to concentrations of 10mM and aliquoted and stored at −80°C. Working concentrations for all chemicals were determined by lethal dose tests.
7
Purine Rescue Agents for Antifolate Drugs
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Pyrimethamine (Sigma-Aldrich), methotrexate (Selleck Chemicals), trimethoprim (Sigma-Aldrich), trimetrexate hydrochloride (CI-898; Santa Cruz) were dissolved in DMSO. Rescue agents were dissolved in water unless indicated otherwise and treatments with hypoxanthine (dissolved in 67% formate; Sigma-Aldrich), folinic acid (5-formyl-THF; Sigma-Aldrich), thymidine (Sigma-Aldrich), glycine (Sigma-Aldrich), adenosine (Sigma-Aldrich), and guanosine (Sigma-Aldrich) were performed as indicated. Compound C1 was synthesized by Specs Compound Handling as described in patent WO2021078995A1.
8
Chemotherapy-Induced T-ALL Cell Apoptosis
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T-ALL cells (10,000 or 40,000 per well) were seeded in 96-well plates and incubated with chemotherapeutic agents at the doses indicated below for 48 h. Chemotherapy doses were as follows unless otherwise indicated: asparaginase, 10 international units/ml (Sigma-Aldrich); dexamethasone, 10 µM (Sigma-Aldrich); vincristine, 1 µM (Selleckchem); doxorubicin, 1 µM (Sigma-Aldrich); etoposide, 10 µM (Sigma-Aldrich); cytarabine, 10 µM (Selleckchem); nelarabine, 10 µM (Sigma-Aldrich); 6-mercaptopurine, 10 µM (Abcam); and methotrexate, 10 µM (Selleckchem). Annexin V and propidium iodide staining were assessed using the Apoptosis Detection kit II (BD Biosciences), and caspase 3/7 activity was assessed using the Caspase Glo 3/7 Assay (Promega) according to the manufacturer’s instructions.
9
Characterization of BRAF-mutant Melanoma Cell Lines
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SK-MEL-28 and 501-mel cells were a gift from Dr. Alfonso Bellacosa at Fox Chase Cancer Center (FCCC). SK-MEL-28 and 501-mel cells were authenticated by the FCCC cell culture core according to ATCC test recommendations. The SK-MEL-28VR1 cell line was identified through progressive vemurafenib selection as previously described [1 (link)]. All cell lines were reanimated less than 6 months before experimentation. Cell lines were cultured in RPMI1640/10%FBS (GenDepot) supplemented with 2mM glutamine (Life Technologies; 25030081) and were maintained at 37C in 5% CO2. Methotrexate, dabrafenib, and encorafenib were obtained from Selleckchem.
10
LPS-Induced Inflammation Modulation
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LPS (Escherichia coli serotype) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Celecoxib, ibrutinib, methotrexate, and ruxolitinib were purchased from Selleck Chemicals (Houston, TX, USA). Primary antibodies against iNOS, TLR4, MyD88, p-TAK1 (Ser412), p-ERK, ERK, p-JNK, p-IKKα/β, IKKα/β, p-NF-κB p65, NF-κB, ASC, caspase-1, and NLRP3 were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-IL-1β antibody was purchased from Novus Biologicals (Centennial, CO, USA). Antibodies against IL-6, COX-1, COX-2, TAK1, p-p38, and p38 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively. Anti-TNF-α antibody was purchased from Abcam (Cambridge, MA, USA). Anti-β-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-mouse IgG-horseradish peroxidase (HRP) and anti-mouse IgG-HRP antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
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