HepG2 cells (JCRB1054) were purchased from the Health Science Research Resources Bank (Japan Health Sciences Foundation, Osaka, Japan) and maintained at 37 °C under an atmosphere of 95% air and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. Cells were immersed in a culture medium containing the indicated concentrations of Zebularine, 5-Aza-2′-deoxycytidine (5-aza-dC), 2-1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)-3-(1H-indol-3-yl) propionic acid (RG108) or PKR inhibitor. Zebularine (Wako Pure Chemical Industries, Osaka, Japan) was dissolved in distilled water as a stock solution. 5-Aza-2′-deoxycytidine, RG108 (Wako Pure Chemical Industries), PKR inhibitor (Merck Millipore, Tokyo, Japan), acetaminophen (Sigma-Aldrich Japan, Tokyo, Japan) and aflatoxin B1 (Wako Pure Chemical Industries) were each dissolved in DMSO as a stock solution.
Zebularine
Manufactured by Fujifilm
Sourced in Japan
Zebularine is a laboratory reagent used in the field of molecular biology and genetics. It is a cytidine analog that can be incorporated into DNA during replication, leading to the inhibition of DNA methylation. Zebularine is primarily employed in research applications to study the role of DNA methylation in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Acetaminophen, by Merck Group (1 mentions)
Rg108, by Fujifilm (1 mentions)
Pkr inhibitor, by Merck Group (1 mentions)
5 aza 2 deoxycytidine, by Fujifilm (1 mentions)
Aflatoxin b1, by Fujifilm (1 mentions)
5 aza 2 deoxycytidine, by Merck Group (1 mentions)
Collagenase type 4, by Merck Group (1 mentions)
5 azacytidine, by Merck Group (1 mentions)
4 protocols using zebularine
1
HepG2 cells epigenetic modulation
2
Zebularine Treatment of Cholangiocarcinoma Cell Lines
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TFK-1 [32 (link)] (RCB2537) and HuCCT1 [33 (link)] (RCB1960) were provided by the RIKEN BRC through the National Bio-Resource Project of MEXT, Japan. KKU-100 (JCRB1568), KKU-M156 (JCRB1561) [34 (link)] and KKU-M213 [35 (link)] (JCRB1557) were provided by the JCRB cell bank at the National Institute of Biomedical Innovation, Japan. TFK-1, HuCCT1 and KKU-M213 were maintained at 37°C under an atmosphere of 95% air and 5% CO2 in RPMI1640 containing 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. KKU-100 and KKU-M156 were maintained at 37°C under an atmosphere of 95% air and 5% CO2 in DMEM containing 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. Cells were immersed in a culture medium containing the indicated Zebularine concentrations. Zebularine (Wako Pure Chemical Industries, Osaka, Japan) was dissolved in distilled water as a stock solution.
3
Epigenetic Modulation Protocol
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The DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine was obtained from Sigma-Aldrich (St. Louis, MO), zebularine was obtained from Fujifilm Wako Pure Chemical Corporation, and the histone deacetylase inhibitor TSA was obtained from Sigma-Aldrich.
4
Modulating TMEM164 expression in human liver cells
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Human liver endothelial cells (TMNK1) and human HCC cells (Huh7) were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). HUVECs were obtained from Lonza Co, Ltd (Tokyo, Japan). Each cell line was cultured according to recommended culture methods. TSA and 5-azacytidine were purchased from Sigma-Aldrich Japan K.K. (Tokyo, Japan). Zebularine was purchased from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan). TMNK1 cells were treated with 75–150 μM TSA, 25–50 μM 5-azacytidine, and 100–200 μM Zebularine, all of which inhibit DNA methylation. TMEM164 expression was evaluated after 24-hour treatment.
To create fresh primary HCV-derived HCC cells, a human HCC sample was suspended in 1 mg/mL type-4 collagenase (Sigma-Aldrich Japan K.K.), red blood cells were removed, and the cells were subcutaneously transplanted into NOD/SCID mice. Subcutaneous tumors were resuspended before use in cell experiments.
To create fresh primary HCV-derived HCC cells, a human HCC sample was suspended in 1 mg/mL type-4 collagenase (Sigma-Aldrich Japan K.K.), red blood cells were removed, and the cells were subcutaneously transplanted into NOD/SCID mice. Subcutaneous tumors were resuspended before use in cell experiments.
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