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Gsk 3484862

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GSK-3484862 is a laboratory reagent produced by MedChemExpress. It is a small molecule compound with a specific chemical structure. The core function of GSK-3484862 is to serve as a tool for scientific research and analysis, without interpretation of its intended use.

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Lab products found in correlation

Quantstudio 3 real time pcr system, by Thermo Fisher Scientific (2 mentions) Anti cd11c, by Thermo Fisher Scientific (1 mentions) Anti sma, by Merck Group (1 mentions) Anti 5 mc, by Cell Signaling Technology (1 mentions) Anti granzyme b, by Cell Signaling Technology (1 mentions) Anti pd l1, by Genentech (1 mentions) Anti erk, by Cell Signaling Technology (1 mentions) Anti dnmt1, by Abcam (1 mentions) Anti phospho erk, by Cell Signaling Technology (1 mentions) Anti f4 80, by BD (1 mentions) Tnf α, by Thermo Fisher Scientific (1 mentions) 5 azacytidine, by Merck Group (1 mentions) Anti gapdh, by Cell Signaling Technology (1 mentions) Egm 2 bulletkit, by Lonza (1 mentions) Anti cd8, by BioLegend (1 mentions) Anti cd4, by BD (1 mentions) Anti cd45, by BD (1 mentions) Ifn γ, by Merck Group (1 mentions) Anti vcam 1, by BD (1 mentions) Fetal bovine serum (fbs), by Sartorius (1 mentions)

6 protocols using gsk 3484862

1

Immunomodulatory Protocols for Cancer Research

Cited 2 times
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Antibodies used in this study were anti-DNMT1 (Abcam), anti-5mC (Cell Signaling), anti-GAPDH, anti-ERK, anti-phospho-ERK (Cell signaling), anti-SMA (Sigma), anti-CD8 (Biolegend, BioXcell, or eBiosciences), anti-CD11c, anti-CD45, anti-F480, anti-MECA-79, anti-Vcam1, and anti-CD4 (all from BD Biosciences), anti-granzyme B (Cell Signaling), anti-mouse IgG (BioXcell), anti-PD-1 (CD279, BioXcell), and anti-PD-L1 (Genentech, MTA program). MGECs were isolated and cultured as described previously by our lab60 ,61 (link). Human umbilical vein endothelial cells (HUVEC) were purchased from Lonza and cultured in EBM-2 supplemented with the EGM-2 bullet kit. Culture dishes were precoated with 1% gelatin. For the labeling of CD8+ T-cells, CellTrackerTM green dye was used (Molecular Probes). The culture of EO771 and PyMT cells was described previously62 (link),63 (link). Other reagents include a JAK2 inhibitor (AG490), NFκΒ inhibitor (JSH23, Sigma), TNFα (Peprotech), IFNγ (EMD Millipore), GSK3484862 (Med Chem Express), and 5-Azacytidine (Sigma).
Kim D.J., Anandh S., Null J.L., Przanowski P., Bhatnagar S., Kumar P., Shelton S.E., Grundy E.E., Chiappinelli K.B., Kamm R.D., Barbie D.A, & Dudley A.C. (2023). Priming a vascular-selective cytokine response permits CD8+ T-cell entry into tumors. Nature Communications, 14, 2122.
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2

GM12878 Cell Culture and GSK-3484862 Treatment

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GM12878 cells were cultured in RPMI (Vivacell) supplied with 10% Fetal bovine serum (Biological Industries) under 5% CO2 at 37°C. For GSK-3484862 (HY-135146, MedChemExpress) treatment, GM12878 cells were plated in 10-cm dishes at a cell density of ∼3 × 106/dish. The next day, cells were treated with 0.1% DMSO or 5 μM GSK-3484862 for four consecutive days. On Day 3 of treatment, the cells were subcultured and DMSO (D2650, Sigma) or GSK-3484862 were replenished.
Xiong X., Chen H., Zhang Q., Liu Y, & Xu C. (2024). Uncovering the roles of DNA hemi-methylation in transcriptional regulation using MspJI-assisted hemi-methylation sequencing. Nucleic Acids Research, 52(5), e24.
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3

DNA Methylation and CTCF Profiling

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The following items were key reagents and kits used in this study: GSK-3484862 (HY-135146, MedChemExpress), NEB T4 ligase (M0202L, NEB), MspJI (R0661S, NEB), EZ DNA Methylation-Lightning Kit (D5030, Zymo), anti-Rabbit IgG antibody (3418S, Cell Signaling Technology), anti-CTCF antibody (2729S, Cell Signaling Technology) and HiFi HotStart Uracil + DNA Polymerase (KK2801, Roche).
Xiong X., Chen H., Zhang Q., Liu Y, & Xu C. (2024). Uncovering the roles of DNA hemi-methylation in transcriptional regulation using MspJI-assisted hemi-methylation sequencing. Nucleic Acids Research, 52(5), e24.
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4

Analyzing KAT6A Regulation by DNMT1 Inhibition

Cited 1 time
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HEK293 cells were transiently transfected with constructs for expression of FLAG-HA-tagged human KAT6A using PEI. Twenty-four hours after transfection, the cells were treated with the DNMT1 inhibitor GSK-3484862 (MedChemExpress, Nr.: HY-135146) at 2 μM and 10 μM for 3 days as well as 0.1% DMSO as a solvent control as described (43 (link)), followed by a chromatin immunoprecipitation using a FLAG antibody and qPCR.
Weber L.M., Jia Y., Stielow B., Gisselbrecht S.S., Cao Y., Ren Y., Rohner I., King J., Rothman E., Fischer S., Simon C., Forné I., Nist A., Stiewe T., Bulyk M.L., Wang Z, & Liefke R. (2022). The histone acetyltransferase KAT6A is recruited to unmethylated CpG islands via a DNA binding winged helix domain. Nucleic Acids Research, 51(2), 574-594.
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5

Selective DNMT1 Inhibition and DNA Methylation Assessment

Cited 3 times
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A selective DNMT1 inhibitor, GSK3484862 (MedChemExpress), was dissolved in dimethylsulfoxide and added to the culture media at a final concentration of 2 μM, based on methods described in previous reports (24 (link), 53 (link)). Forty-eight hours later, GSK3484862 was removed from the media by replacing it with fresh culture media.
Methylation quantification by MSRE-qPCR was performed as we previously described (23 (link)). Briefly, 150 ng genomic DNA was digested with 10 units of methylation-sensitive HpaII (New England BioLabs) at 37°C for 2 hours. Relative amounts of digested samples and serially diluted undigested samples were measured by qPCR with KOD SYBR (TOYOBO) using the QuantStudio 3 Real-time PCR system (Thermo Fisher Scientific). In every sample, a melt curve with a single peak was confirmed. Methylation levels were calculated on the basis of Ct values for digested samples, and the calibration curves were generated from Ct values for serially diluted undigested samples. Primer sequences are listed in Supplemental Table 3.
Iwasaki Y., Aksu C., Reyes M., Ay B., He Q, & Bastepe M. (2023). The long-range interaction between two GNAS imprinting control regions delineates pseudohypoparathyroidism type 1B pathogenesis. The Journal of Clinical Investigation, 133(8), e167953.
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6

Evaluating Novel Chemical Compounds

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Chemicals used in this study include: GSK-3484862 (MedChemExpress, HY-135146), GSK-3685032 (MedChemExpress, HY-139664), Decitabine (DAC, 5-Aza-2′-deoxycytidine, Millipore Sigma, A3656), MG132 (Millipore Sigma, 474787), and Aphidicolin (Cayman Chemicals, 14007).
Chen Q., Liu B., Zeng Y., Hwang J.W., Dai N., Corrêa IR J.r., Estecio M.R., Zhang X., Santos M.A., Chen T, & Cheng X. (2023). GSK-3484862 targets DNMT1 for degradation in cells. NAR Cancer, 5(2), zcad022.
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