Zebularine, RG108 were purchased from Cayman Chemical. Decitabine,FCDR and EGCG were purchased from Sigma-Aldrich.
Rg108
Manufactured by Cayman Chemical
Sourced in United States
RG108 is a chemical compound used for research and development purposes. It serves as a selective and reversible inhibitor of different cellular processes. The core function of RG108 is to modulate the activity of target biomolecules in controlled experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Zebularine, by Cayman Chemical (1 mentions)
Decitabine, by Merck Group (1 mentions)
Epigallocatechin gallate (egcg), by Merck Group (1 mentions)
High fat diet (hfd), by Research Diets (1 mentions)
Ex 527, by Cayman Chemical (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
N acetyl l cysteine nac, by Merck Group (1 mentions)
Pen strep, by Merck Group (1 mentions)
Carbonyl cyanide 3 chlorophenylhydrazone cccp, by Merck Group (1 mentions)
Dnmt activity inhibitor assay kit, by Epigentek (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Pcdna3 myc dnmt3b1, by Addgene (1 mentions)
Pcdna3 myc dnmt3a, by Addgene (1 mentions)
Pcdna3 myc dnmt1, by Addgene (1 mentions)
5 protocols using rg108
1
Epigenetic Modulators in Drug Discovery
2
Investigating TonEBP in Obese Mice
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All the methods involving live mice were carried out in accordance with the approved guidelines. All experimental protocols were approved by Institutional Animal Care and Use Committee of the Ulsan National Institute of Science and Technology (UNISTACUC-12-15-A).
All studies used male C57BL/6 J background mice. TonEBP haplo-deficient mice (TonEBP + /Δ) were described previously5 (link). Mice carrying the loxP-targeted TonEBP gene (TonEBPfl/fl) were provided by Dr Neuhofer48 (link). Leptin receptor mutant db/db (Leprdb/db) mice and Adiponectin-Cre transgenic mice (AQ-cre) were obtained from The Jackson Laboratory (USA). TonEBPfl/fl mice were bred with AQ-Cre knock-in mice to generate TonEBPfl/flAQ-Cre mice.
For obese mice model, mice at 8 weeks age were fed either a CD (10% fat as kcal, Research Diets, NJ, USA) or HFD (60% fat as kcal, Research Diets) during 16 weeks. Mice were intraperitoneally injected with RG108 (Cayman, MI, USA) at 12 mg/kg every 2 days.
All studies used male C57BL/6 J background mice. TonEBP haplo-deficient mice (TonEBP + /Δ) were described previously5 (link). Mice carrying the loxP-targeted TonEBP gene (TonEBPfl/fl) were provided by Dr Neuhofer48 (link). Leptin receptor mutant db/db (Leprdb/db) mice and Adiponectin-Cre transgenic mice (AQ-cre) were obtained from The Jackson Laboratory (USA). TonEBPfl/fl mice were bred with AQ-Cre knock-in mice to generate TonEBPfl/flAQ-Cre mice.
For obese mice model, mice at 8 weeks age were fed either a CD (10% fat as kcal, Research Diets, NJ, USA) or HFD (60% fat as kcal, Research Diets) during 16 weeks. Mice were intraperitoneally injected with RG108 (Cayman, MI, USA) at 12 mg/kg every 2 days.
3
Epigenetic Modulation of cES Cells
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cES cells were obtained by culturing ES cells in conditioning medium (CM) consisting of BGM supplemented with 5 μM Ex527 and 100 μM RG108 (Cayman chemical, 10009798 and 13302, respectively) for 10 cell passages. The cells were passaged 1:15–1:20 every 2 days. Ex527 and RG108 were added from stock solutions in dimethyl sulfoxide (DMSO). The total DMSO concentration in CM and BGM was 0.03% (v/v).
4
Oxidative Stress and Cardiac Fibroblasts
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Human cardiac fibroblasts (HCFs) (Innoprot) were grown as previously described77 (link). HCFs were exposed 24 h to the following chemicals to induce oxidative stress: 200 µM H2O2 (Carl Roth) or 10 µM Carbonyl cyanide 3-chlorophenylhydrazone (CCCP; Sigma-Aldrich). 10mM N-Acetyl-L-cysteine (NAC; Sigma-Aldrich) was added to HCF medium 16 h prior H2O2 or CCCP treatment as ROS scavanger. 50 µM of RG108 (Cayman Chemical) was added to HCF medium as DNMT inhibitor. Differentiated H9C2 rat cardiomyoblasts (RRID: CVCL_0286), derived from the embryonic left ventricle an E13 BDIX female rat heart, were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 1% donor calf serum (DCS - Sigma), 1% Pen-Strep (Sigma) and 1% Glutamine (Gibco) to achieve cardiac differentiation.
5
DNMT Isoform Selectivity Assay
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Compounds 1, 22 and 24 were tested to evaluate their selectivity on the different DNMT isoforms. To work selectively on DNMT1, DNMT3A, and DNMT3B, HEK293T cells were transfected with the plasmids containing the three different DNMTs' sequences and mock control. Cells were transfected with 2.5 μg of expression plasmid using Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, USA) according the manufacturer's instruction. Plasmids pcDNA3/Myc-DNMT1 (Addgene plasmid # 36939), pcDNA3/Myc-DNMT3A (Addgene plasmid # 35521), and pcDNA3/Myc-DNMT3B1 (Addgene plasmid # 35522) were a gift from Arthur Riggs. [39, 40] The presence of exogenous DNMTs was checked by Western blot (not shown), and afterward, the transfected cells were freshly lysed in RIPA buffer as above. Cellular extract of 35 μg were incubated with selected compounds at different concentration in the 1-150 μm range (1% DMSO final conc.) or with vehicle alone (1% DMSO) at 37 °C for 2 h. RG108 (Cayman) was used as positive controls, while as negative control, lysates were denatured at 100 °C for 30 min. DNMT activity was detected by DNMT activity/inhibitor assay kit (Epigentek). Data are presented as means ± SD; each compound was tested at least three times.
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