H358, ER-HRASG12V MCF10A and KP (tetON) cells were plated in 96 well plates and the following day co-transfected with pRL-TK control and pGL3-3′UTR PD-L1 luciferase constructs using Lipofectamine 2000 (Life Technologies). 24 h after transfection, PMA (200 nM; Sigma), doxycycline (1 μg/ml; Sigma) or MEK inhibitor GSK1120212 (25 nM; Selleckchem) was added, and 6-7 h later the Dual-Luciferase Reporter Assay (Promega) was performed. For ER-HRASG12V MCF10A, 24 h after transfection cells were serum-starved overnight, and then treated with 4-OHT (100 nM) for 24 h before the Dual-Luciferase Reporter Assay (Promega) was performed.
Gsk1120212
Manufactured by Selleck Chemicals
Sourced in United States, China, Germany
The GSK1120212 is a laboratory equipment product. It is a high-performance liquid chromatography (HPLC) instrument used for the separation, identification, and quantification of chemical compounds in complex mixtures. The GSK1120212 employs advanced chromatographic techniques to provide accurate and reliable analytical results for various research and development applications.
Automatically generated - may contain errors
Lab products found in correlation
Plx4032, by Selleck Chemicals (10 mentions)
Gsk2118436, by Selleck Chemicals (7 mentions)
Pd0325901, by Selleck Chemicals (6 mentions)
Azd6244, by Selleck Chemicals (5 mentions)
Sch772984, by Selleck Chemicals (4 mentions)
Bkm120, by Selleck Chemicals (4 mentions)
Plx4720, by Selleck Chemicals (4 mentions)
Trametinib, by Selleck Chemicals (4 mentions)
Mk 2206, by Selleck Chemicals (3 mentions)
Bez235, by Selleck Chemicals (3 mentions)
Ly294002, by Selleck Chemicals (3 mentions)
Gdc 0973, by Selleck Chemicals (3 mentions)
Ly3214996, by Selleck Chemicals (3 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Mg132, by Selleck Chemicals (2 mentions)
Lapatinib, by Selleck Chemicals (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Erk1 2, by Cell Signaling Technology (2 mentions)
Bay80 6946, by Selleck Chemicals (2 mentions)
51 protocols using gsk1120212
1
Regulation of PD-L1 3'UTR Activity
2
Targeting RICTOR and KRAS in Cancer
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Cells were obtained from American Type Tissue Collection (Manassas, VA) or collaborators, authenticated via STR DNA fingerprinting at UT MD Anderson Characterized Cell Line Core. Whole genome SNP array profiling was used to determine RICTOR amplified (copy number variation (CNV ≥ 3.5) and non-amplified cell lines (CNV ~2) [31 (link)]. Immortalized human bronchial epithelial cells expressing wtKRAS (HBEC3-KT) and KRAS mutant with stable p53 knockdown (HBEC3-KT53KC12) lines were provided by Drs. Gazdar and Minna (UT Southwestern Medical Center, Dallas, TX). Stable RICTOR knockdown was developed using pTRIPZ inducible lentiviral shRNA plasmids (Rictor shRNA #RHS4696, Non-silencing shRNAmir Control (NTC) #RHS4743) (GE Dharmacon, Lafayette, CO). Targeted inhibitors AZD2014 (vistusertib), AZD6244 (selumetinib), and GSK1120212 (trametinib) were obtained from Selleck Chemicals (Houston, TX).
3
Selective Kinase Inhibitor Protocol
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VS-5584, GSK1120212, SCH772984, and BVD-523 were purchased from Selleck Chemicals (Houston, TX, USA).
4
Cell Line Maintenance and Reagents
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AU565, HCC1419, NCI-H2170, HCC202, HCC1954, NCI-N87, ZR75-1, SKOV3, ZR75-30, MDAMB175VII, CALU3, MDAMB453, MDAMB361, JIMT1, SKBR3 and HCC2218 cells were obtained from ATCC. OE19 and OE33 were obtained from ECCC; COLO-678 was obtained from DSMZ, and KYSE-410 from Sigma-Aldrich. BT-474-M3 cells (hereafter simply referred to as BT-474) were obtained from Hermes biosciences. All cell lines were maintained in RPMI supplemented with 10% FBS, penicillin, and streptomycin. GSK-1120212 and MK-2206 were purchased from Selleckchem. Recombinant human HRG-β1 (EGF domain) was from R&D Systems.
5
Inhibitor Treatment of Cultured Cells
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BEZ235 (dual PI3K/mTOR inhibitor), BKM120 (PI3K inhibitor), everolimus (mTOR inhibitor) and GSK1120212 (MEK inhibitor) were purchased from Selleck Chemicals (Houston, TX, USA) and were prepared as 2.1 mmol/L, 24 mmol/L, 10 mmol/L and 14 mmol/L stock solutions, respectively, in DMSO and stored at −80°C. Cultured cells were treated for 72 hours with a medium changes and fresh drug additions as indicated in the figure legends.
6
Establishing Melanoma Resistance Models
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Melanoma cell lines and HEK293T cells were cultured in DMEM containing fetal bovine serum (FBS) (Sigma), 2 mM glutamine, 100 U/ml penicillin, and 0.1 mg/ml streptomycin (all Gibco) under standard conditions. Resistant cell lines were generated by treatment with increasing concentrations of PLX4720 (Selleck Chemicals, up to 3 μM) or the combination of dabrafenib (GSK2118436, Abmole, up to 0.5 μM)/trametinib (GSK1120212, S2673, Selleck Chemicals, up to 50 nM). Transfections and production of lentivirus were performed as described previously (Vredeveld et al., 2012 (link)). A375R cells were infected and selected with puromycin (1 μg/ml). For the colony formation assays, 20,000 cells were seeded in six-well plates and indicated concentrations of PLX4720, vemurafenib, or LY3009120 were added the next day. Cells were stained by 0.1% crystal violet in 50% methanol and 50% H20. After staining, de-staining by 10% acetic acid was used to quantify the number of stained cells. Color intensity was measured at 590 nm and values were normalized to DMSO control. For immunoblotting, cells were treated for 24 hr with the indicated concentrations of BRAFi.
7
Anticancer Inhibitor Compound Preparation
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The HDACi (LBH589, SAHA and FK228) were purchased from Selleck Chemicals, Munich, Germany. The MAPK inhibitors, GSK1120212 and ARRY-162, were obtained from Selleck Chemicals and AbMole BioScience, HongKong, China, respectively. FAK inhibitor TAE226 was from Selleck Chemicals and PF573228 was from Tocris Bioscience, Bristol, UK. Phosphatidylinositol 3-kinases (PI3K) inhibitors GDC-0941 and BKM120 were obtained from Selleck Chemicals. All compounds were dissolved in dimethyl sulfoxide and stock drug solutions were diluted in complete RPMI-1640 culture medium (Life Technologies, Mulgrave, VIC, Australia) to various concentrations for experimentation.
8
Establishing ALK-dependent Cell Lines
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Cell lines used, with their respective mutations, are listed in Supplementary Table S1 . The murine interleukin 3 (Il-3)-dependent pro-B-Cell line, Ba/F3, was used as a model to obtain ALK-dependent cells, owing to its potent growth capacity and use of kinase oncogene signalling53 (link). Cells were grown in RPMI 1640 (Invitrogen) supplemented with 10% FCS, 100IU/ml penicillin/streptomycin, 2 mM L-glutamine and 25 mM HEPES (Life Technologies). Ba/F3 cells were cultured in the presence of 1 ng/mL Il-3 (213-13, Peprotech). Cells were cultured at 37 °C in a 5% CO2/95% O2 humidified environment. Mycoplasma testing (Lonza) and short tandem repeat genotyping were regularly performed.
Cells were treated for indicated time periods with inhibitory compounds: the ALK inhibitors, NVP-TAE-684 (0.32 µM, S1108, Novartis/SelleckChem), crizotinib (0.5 µM, PF-02341066, S1068, Pfizer/SelleckChem) or LDK-378 (0.2 µM, S7083, SelleckChem); the MEK inhibitors, U-0126 (8 µM, Sigma-Aldrich) or trametinib (0.05 µM, GSK1120212, S2673, SelleckChem); the dual PI3K/mTOR inhibitor, NVP-BEZ-235 (0.5 µM, S1009_3, SelleckChem) or the PI3K inhibitor pictilisib (0.5 µM, S1065, SelleckChem). Dimethyl sulfoxide (DMSO, VWR) was used to dissolve the compounds and served as vehicle control.
Cells were treated for indicated time periods with inhibitory compounds: the ALK inhibitors, NVP-TAE-684 (0.32 µM, S1108, Novartis/SelleckChem), crizotinib (0.5 µM, PF-02341066, S1068, Pfizer/SelleckChem) or LDK-378 (0.2 µM, S7083, SelleckChem); the MEK inhibitors, U-0126 (8 µM, Sigma-Aldrich) or trametinib (0.05 µM, GSK1120212, S2673, SelleckChem); the dual PI3K/mTOR inhibitor, NVP-BEZ-235 (0.5 µM, S1009_3, SelleckChem) or the PI3K inhibitor pictilisib (0.5 µM, S1065, SelleckChem). Dimethyl sulfoxide (DMSO, VWR) was used to dissolve the compounds and served as vehicle control.
9
Therapeutic Compound Reconstitution and Preparation
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MEK inhibitor GSK1120212/trametinib and GSK211436/dabrafenib were purchased from Selleck Chemicals (Houston, TX, USA). HSP90 inhibitors used were ganetespib (STA-9090, MedChemExpress) and 17-AAG (Bioconnect). The metabolic poison phenyl arsine oxide (PAO) and solvent dimethylsulfoxide were obtained from Sigma-Aldrich (St Louis, MO, USA). Dacarbazine was obtained from the Slotervaart hospital pharmacy. AG-879 was bought from MedChemExpress (HY-20878). All drugs except dacarbazine (which was already dissolved) were reconstituted in 100% dimethylsulfoxide to a final concentration of 1–10 mM. Recombinant human TNF (Peprotech) and IFNγ (R&D Systems) were diluted in sterile H2O to a final concentration of 100–200 μg/mL. Peptides for MART-1, CDK4, and NY-ESO-1 were reconstituted in 100% dimethylsulfoxide to a final concentration of 10 mM.
10
Murine Pancreatic Cancer Induction and Treatment
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To induce pancreatitis, mice aged 4–6 weeks were treated with 8-hourly intraperitoneal (i.p.) injections of caerulein (Sigma-Aldrich) at a dosage of 75 µg/kg body weight over 2 consecutive days. DT (Enzo Life Science) was administered i.p. every 4 days at a concentration of 25 ng/g. To establish the subcutaneous tumour model, 2×106 of primary mouse pancreatic cancer cell lines iKras*1, iKras*2 and iKras*3 derived from iKras*p53* tumours,11 (link)
13 (link) 4×105 of 65 671 cells (FVB/NJ strain)14 (link) or 7940B cells (C57BL/6J strain)15 (link) derived from KPC tumour (P48-Cre; loxP-stop-loxP (LSL)-KrasG12D; p53flox/+) were injected into CD11d-DTR mice of compatible genetic background. Tumour diameters were measured with digital callipers and the tumour volume was calculated by the formula: length×width2/2. For CD8+ T-cell depletion, anti-CD8 monoclonal antibody (BioXcell #BE0061; clone 2.43; 200 µg/mouse) was injected i.p. twice per week. Purified anti-mPD-1 antibody (BioXcell #BE0033-2; clone J43) was used for in vivo PD-1 blockade at a dose of 200 μg/i.p. injection, repeated every 3 days if needed. MEK inhibitor (MEKi) GSK1120212 (Selleckchem) was administered daily (1 mg/kg, i.p.) in a 10% (2-Hydroxypropyl)-β-cyclodextrin solution.
13 (link) 4×105 of 65 671 cells (FVB/NJ strain)14 (link) or 7940B cells (C57BL/6J strain)15 (link) derived from KPC tumour (P48-Cre; loxP-stop-loxP (LSL)-KrasG12D; p53flox/+) were injected into CD11d-DTR mice of compatible genetic background. Tumour diameters were measured with digital callipers and the tumour volume was calculated by the formula: length×width2/2. For CD8+ T-cell depletion, anti-CD8 monoclonal antibody (BioXcell #BE0061; clone 2.43; 200 µg/mouse) was injected i.p. twice per week. Purified anti-mPD-1 antibody (BioXcell #BE0033-2; clone J43) was used for in vivo PD-1 blockade at a dose of 200 μg/i.p. injection, repeated every 3 days if needed. MEK inhibitor (MEKi) GSK1120212 (Selleckchem) was administered daily (1 mg/kg, i.p.) in a 10% (2-Hydroxypropyl)-β-cyclodextrin solution.
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