Foreskin fibroblast cells (BJ and IMR90) obtained from ATCC were cultured in Dulbecco Modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS). hTERT-HK used in our previous study [28 ] were grown in FAD medium, containing Ham's F-12 medium (Gibco), Dulbecco Modified Eagle medium (Gibco), 2% fetal calf serum, adenine (Sigma), 10 ng/ml human epidermal growth factor (STEMCELL technologies), 5 μg/ml insulin (Sigma), and 400 μg/ml hydrocortisone (Sigma). HPV(+)SCC cell lines UD-SCC-2, and VU-SCC-147 were kindly provided by Dr. Robert Ferris and cultured in 10% DMEM. MCV(+)MCC cell lines MKL-1, MKL-2, MS-1, and CVG-1 were cultured in RPMI medium supplemented with 10% FCS and non-essential amino acids. 293 FT cells (Invitrogen) for lentivirus production were cultured in 10% FCS DMEM. EZH2 degraders MS177 and MS1943 were purchased from MedChemExpress, and EZH2-specific methyltransferase inhibitor GSK126 was purchased from Selleckchem. Tazemetostat was kindly provided by Dr. Jan Beumer.
Ms1943
Manufactured by MedChemExpress
Sourced in United States
MS1943 is a laboratory equipment product from MedChemExpress. It is a precision instrument used for mass spectrometry analysis, a widely-used analytical technique in various scientific fields, including chemistry, biochemistry, and life sciences.
Automatically generated - may contain errors
Lab products found in correlation
Gsk126, by Selleck Chemicals (2 mentions)
Human epidermal growth factor (hegf), by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Ms 177, by MedChemExpress (1 mentions)
Ham s f12 medium, by Thermo Fisher Scientific (1 mentions)
293ft cell, by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Adenine, by Merck Group (1 mentions)
Ibrutinib, by MedChemExpress (1 mentions)
Tazemetostat, by MedChemExpress (1 mentions)
Lapatinib, by MedChemExpress (1 mentions)
Ribavirin, by Selleck Chemicals (1 mentions)
Dznep, by Merck Group (1 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
Epz 6438, by Selleck Chemicals (1 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Quikchange 2 site directed mutagenesis kit, by Agilent Technologies (1 mentions)
4 protocols using ms1943
1
Culturing Diverse Cell Lines for EZH2 Studies
2
EZH2 Inhibitor and Degrader Protocols
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Tazemetostat (iEZH2, EZH2 inhibitor), MS1943 (dEZH2, EZH2 degrader), and Ibrutinib (Ib, BTK inhibitor) were purchased from MedChemExpress (NJ, USA). These drugs were dissolved in dimethyl sulfoxide (DMSO) as recommended by the manufacturer and stored at -80°C. All these drugs were diluted with the RPMI 1640 medium with 10% heat-inactivated FBS, 2mM L-glutamine and 1% antibiotics before used for the treatment of cell lines.
3
Preparation and Application of Targeted Inhibitors
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Lapatinib (HER2/neu-EGFR inhibitor) and MS1943 (EZH2 degrader) were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Both were dissolved in dimethyl sulfoxide (DMSO) as recommended by the manufacturer and stored at −80 °C. All these drugs were diluted with the RPMI 1640 medium with 10% heat-inactivated FBS, 2 mM L-glutamine, and 1% antibiotics before being used for the treatment of the cell lines.
4
Epigenetic Modulation of EZH2 Function
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The chemicals used for treating cells were GSK126 (Selleckchem, S7061), EPZ6438 (Selleckchem, S7128), Sappanone A (Cayman Chemicals, 23205), MPA (Selleckchem, S2487), Ribavirin (Selleckchem, S2504), DZNep (Sigma, S804983) and MS1943 (MedChemExpress, HY-133129); all are listed in Table S1. S1. pCMVHA hEZH2 and V5-EZH2 vector was used to generate EZH2-H689A mutant vector using the mutagenesis primers listed in Table S1 with QuikChange II site-directed mutagenesis kit (Agilent) following the manufacturer's instructions. Custom designed siRNA oligonucleotides listed in Table S1 were purchased from Bioneer Pacific. For transient transfection, 25x10 4 cells were transfected with 2.5 µg of DNA using Lipofectamine 3000 transfection reagent (Invitrogen). For siRNA experiments, 25x10 4 cells were transfected with 10 nM of the indicated oligonucleotides in Table S1 using the Lipofectamine RNAiMAX transfection reagent (Invitrogen). 72 hours after siRNA transfection, cells were used for functional assays or collected for western blot analysis.
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