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Dznep

Manufactured by MedChemExpress
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DZNep is a chemical compound used as a research tool. It functions as an inhibitor of the histone methyltransferase EED, which is part of the Polycomb Repressive Complex 2 (PRC2). DZNep is often utilized in scientific research to study the role of epigenetic regulation mediated by the PRC2 complex.

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Lab products found in correlation

Celltiter glo luminescent cell viability assay kit, by Promega (2 mentions) Celltiter glo luminescent cell viability assay, by Promega (2 mentions) Epz6438, by MedChemExpress (2 mentions) Hepg2, by BNCC (2 mentions) Hep3b, by BNCC (2 mentions) 5 aza 2 deoxycytidine, by Merck Group (2 mentions) Lo2 cell line, by Procell (2 mentions) Gsk126, by MedChemExpress (1 mentions) Anti cd14 magnetic bead, by Miltenyi Biotec (1 mentions) Zebularine, by Merck Group (1 mentions) Mnase, by New England Biolabs (1 mentions) Wheat germ agglutinin (wga), by Merck Group (1 mentions) Haeiii, by New England Biolabs (1 mentions) Methylstat, by Merck Group (1 mentions) Fitc dextran, by Merck Group (1 mentions) Benzonase, by Merck Group (1 mentions) Ecori, by New England Biolabs (1 mentions) Xhoi, by New England Biolabs (1 mentions) Defactinib, by MedChemExpress (1 mentions) Losartan, by MedChemExpress (1 mentions)

7 protocols using dznep

1

Hepatoma Cell Culture and Treatment Protocols

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The human hepatoma cell lines PLC/PRF/5, Huh7, and Hep3B used in this study were purchased from the American Type Culture Collection (Manassas, VA, USA). PLC/PRF/5 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium, and Huh7 and Hep3B cell lines were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum at 37 °C and 5% CO2. Hepatoma cells were treated with recombinant interferon gamma (IFNγ) (Sino Biological Inc.), DZNep (MedChemExpress, Monmouth Junction, NJ, USA), or GSK-126 (MedChemExpress) for different times and at different concentrations.
Monocytes were selected from peripheral blood mononuclear cells using anti-CD14 magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany) as described previously [33 (link)].
Xiao G., Jin L.L., Liu C.Q., Wang Y.C., Meng Y.M., Zhou Z.G., Chen J., Yu X.J., Zhang Y.J., Xu J, & Zheng L. (2019). EZH2 negatively regulates PD-L1 expression in hepatocellular carcinoma. Journal for Immunotherapy of Cancer, 7, 300.
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2

Evaluating Cell Viability under Stress

Cited 2 times
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Cell viability was measured by CellTiter-Glo Luminescent Cell Viability Assay (Promega) according to manufacturer’s instructions after exposing 3 days under different conditions including low-glucose (0.45 g/l glucose)10 (link), hypoxia (1% oxygen)7 (link), BMI1 inhibitor (PTC596, PTC therapeutics; PTC209, PTC therapeutics), and EZH2 inhibitor (EPZ6438, Medchemexpress LLC; DZNep, SELLECKCHEM). 1×106 cells were seeded for cell viability under low-glucose or/and hypoxia conditions, 2×103 cells were seeded for drug response under BMI1 or EZH2 inhibitor treatment. IC50 values of BMI1 or EZH2 inhibitors were calculated with GraphPad Prism software. For competitive response assay of BMI1 and EZH2 inhibitors, the proportion of GFP or mCherry positive cells was determined by flow cytometry after treatment the co-cultured MES83-GFP and PN528-mCherry cells with PTC-209, and DZNep. Two-dimensional titration assay was performed on four different subtype GSCs after combined treatment with different doses of PTC596 and EPZ6438. Relative cell growth was measured over a four-day time course using CellTiter-Glo Luminescent Cell Viability Assay kit (Promega) after seeding 1×103 cells.
Jin X., Kim L.J., Wu Q., Wallace L.C., Prager B.C., Sanvoranart T., Gimple R.C., Wang X., Mack S.C., Miller T.E., Huang P., Valentim C.L., Zhou Q.G., Barnholtz-Sloan J.S., Bao S., Sloan A.E, & Rich J.N. (2017). Targeting Glioma Stem Cells through Combined BMI1 and EZH2 Inhibition. Nature medicine, 23(11), 1352-1361.
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3

Xenopus Egg Extract Nuclear Manipulation

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DZNep (MedChemExpress, HY-12186), Methylstat (Sigma, SML0343-5MG), VPA (MedChemExpress, HY-10585), or NDMA (Sigma, N7756) was added to X. laevis egg extract after nuclear assembly. Zebularine (Sigma, Z4775) was added prior to nuclear assembly. For digesting nuclear DNA, 2.5 U/µL Benzonase (Sigma, E1014), 40 U/µL MNase (NEB, M0247S), 0.2 U/μL EcoRI (NEB, R0101S), 0.2 U/μL XhoI (NEB, R0101S), or 0.5 U/μL HaeIII (NEB, R0108S) was added to X. laevis egg extract after nuclear assembly. WGA (Sigma, L9640) and 150-kDa FITC-dextran (Sigma, 69658) were used where indicated.
Chen P., Mishra S, & Levy D.L. (2023). Nuclear growth and import can be uncoupled. bioRxiv.
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4

Evaluating Cell Viability under Stress

Cited 2 times
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Cell viability was measured by CellTiter-Glo Luminescent Cell Viability Assay (Promega) according to manufacturer’s instructions after exposing 3 days under different conditions including low-glucose (0.45 g/l glucose)10 (link), hypoxia (1% oxygen)7 (link), BMI1 inhibitor (PTC596, PTC therapeutics; PTC209, PTC therapeutics), and EZH2 inhibitor (EPZ6438, Medchemexpress LLC; DZNep, SELLECKCHEM). 1×106 cells were seeded for cell viability under low-glucose or/and hypoxia conditions, 2×103 cells were seeded for drug response under BMI1 or EZH2 inhibitor treatment. IC50 values of BMI1 or EZH2 inhibitors were calculated with GraphPad Prism software. For competitive response assay of BMI1 and EZH2 inhibitors, the proportion of GFP or mCherry positive cells was determined by flow cytometry after treatment the co-cultured MES83-GFP and PN528-mCherry cells with PTC-209, and DZNep. Two-dimensional titration assay was performed on four different subtype GSCs after combined treatment with different doses of PTC596 and EPZ6438. Relative cell growth was measured over a four-day time course using CellTiter-Glo Luminescent Cell Viability Assay kit (Promega) after seeding 1×103 cells.
Jin X., Kim L.J., Wu Q., Wallace L.C., Prager B.C., Sanvoranart T., Gimple R.C., Wang X., Mack S.C., Miller T.E., Huang P., Valentim C.L., Zhou Q.G., Barnholtz-Sloan J.S., Bao S., Sloan A.E, & Rich J.N. (2017). Targeting Glioma Stem Cells through Combined BMI1 and EZH2 Inhibition. Nature medicine, 23(11), 1352-1361.
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5

Epigenetic Regulation in Liver Cancer

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The human HCC-derived Hep3B, Huh-7 and HepG2 were obtained from the BeNa Culture Collection (Shanghai, China). LO2 cell line was acquired from Procell Life Technology (Wuhuan, China). All cell lines were authenticated using short tandem repeat (STR) profiling at the time of purchase. Low-passage cells were used for experiments within a period of 6 months after resuscitation. All cells were negative for mycoplasma contamination. All cell lines were cultivated cultured routinely in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum at 37 °C in a humidified 5% CO2 incubator. DZNep was purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA) and dissolved in ddH2O. 5-aza-2′-deoxycytidine was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in DMSO. For the 5-Aza-dC treatment, 5-Aza-dC (5 μM) was replenished daily for 72 h. For the DZNep treatment, DZNep (10 μM) was added to the culture medium for 72 h.
Chen W., Tang D., Tang D, & Dai Y. (2020). Epigenetic silencing of ZIC4 contributes to cancer progression in hepatocellular carcinoma. Cell Death & Disease, 11(10), 906.
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6

Xenograft Tumor Model in BALB/C Mice

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For animal experiments, female, 4-to 6-week-old BALB/C nude mice were purchased from Beijing HFK Bioscience Co., Ltd. (Beijing, China), and were maintained according to protocols approved by the Ethical Committee on Animal Experiments of the Animal Care Committee of Wuhan University (S07921060J). For co-implanted assay, the sphere forming cells and fibroblasts were resuspended in a PBS/Matrigel (BD Biosciences) mixture (1:1 volume), which were then injected in the into the subcutaneous tissue of mouse flanks using 27-gauge needles. For drug administration assay, the Losartan, Defactinib, and DZNep, all purchased from MedChemExpress (MCE, USA), were injected intraperitoneally into the BALB/C nude mice bearing with co-implanted xenograft.
Zhao H., Li R., Chen Y., Yang X, & Shang Z. (2023). Stromal nicotinamide N-methyltransferase orchestrates the crosstalk between fibroblasts and tumour cells in oral squamous cell carcinoma: evidence from patient-derived assembled organoids. Oncogene, 42(15).
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7

Epigenetic Modulation in Liver Cancer

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The human HCC-derived Hep3B, Huh-7 and HepG2 were obtained from the BeNa Culture Collection (Shanghai, China). LO2 cell line was acquired from Procell Life Technology (Wuhuan, China). All cell lines were cultivated cultured routinely in Dulbecco's modifed Eagle's medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum at 37℃ in a humidi ed 5% CO 2 incubator. DZNep was purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA) and dissolved in ddH 2 O. 5-aza-2'deoxycytidine was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in DMSO. For the 5-Aza-dC treatment, 5-Aza-dC (5 µM) was replenished daily for 72 h. For the DZNep treatment, DZNep (10 µM) was added to the culture medium for 72 h.
Wang J., Yang Z., Long F., Luo L., Gao L., Chen C., Sun D., & Tang D. (2020). EZH2-mediated epigenetic silencing of ZIC4 in hepatocellular carcinoma.
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