HNSCC cell lines were kindly gifted as follows: UM-SCC47 from Dr. Thomas Carey (University of Michigan); TU167 from Dr. Jeffrey Myers, (UT MD Anderson Cancer Center). Cell lines were verified by short tandem repeat analysis and were confirmed negative for mycoplasma infection. The p16+/HPV+ (SCC47) and p16−/HPV− (TU167) human HNSCC tumor cells were grown in DMEM (ThermoFisher, Waltham, MA) supplemented with 10% FBS and 1% penicillin-streptomycin. Cells were treated with 50µM MK2 inhibitor (PF 3644022, Tocris), RT (RT; 10Gy), both MK2i and RT, or nothing (control). Cells were irradiated using the MultiRad 225 irradiator. MK2i in low serum media (0.5% FBS) was applied to cells 4 hours prior to irradiation; immediately following RT media was exchanged for media containing 10% FBS and MK2i as previously described [33 (link)]. Cells were grown for an additional 48 hours at which time they were harvested for protein and mRNA analysis. These initial cell experiments were replicated with additional cell lines (SCC25, CAL27, FaDU) which were obtained from ATCC.
Mk2 inhibitor pf 3644022
Manufactured by Bio-Techne
MK2 inhibitor (PF 3644022) is a small molecule compound that inhibits the activity of the Mitogen-Activated Protein Kinase-Activated Protein Kinase 2 (MAPKAPK2 or MK2) enzyme. MK2 is involved in various cellular processes, including inflammation and stress response.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Cal27, by American Type Culture Collection (2 mentions)
Scc25, by American Type Culture Collection (2 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Tween 20, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Plus reagent, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Flag peptide, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Proteinase k, by Thermo Fisher Scientific (1 mentions)
Rnase a, by Thermo Fisher Scientific (1 mentions)
Isopropanol, by Merck Group (1 mentions)
Lipofectamine, by Thermo Fisher Scientific (1 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
Puromycin, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Jackson ImmunoResearch (1 mentions)
Superscript 3 first strand synthesis system, by Thermo Fisher Scientific (1 mentions)
3 protocols using mk2 inhibitor pf 3644022
1
HNSCC Cell Lines Treated with MK2 Inhibitor
2
HNSCC Cell Lines Treated with MK2 Inhibitor
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HNSCC cell lines were kindly gifted as follows: UM-SCC47 from Dr. Thomas Carey (University of Michigan); TU167 from Dr. Jeffrey Myers, (UT MD Anderson Cancer Center). Cell lines were verified by short tandem repeat analysis and were confirmed negative for mycoplasma infection. The p16+/HPV+ (SCC47) and p16−/HPV− (TU167) human HNSCC tumor cells were grown in DMEM (ThermoFisher, Waltham, MA) supplemented with 10% FBS and 1% penicillin-streptomycin. Cells were treated with 50µM MK2 inhibitor (PF 3644022, Tocris), RT (RT; 10Gy), both MK2i and RT, or nothing (control). Cells were irradiated using the MultiRad 225 irradiator. MK2i in low serum media (0.5% FBS) was applied to cells 4 hours prior to irradiation; immediately following RT media was exchanged for media containing 10% FBS and MK2i as previously described [33 (link)]. Cells were grown for an additional 48 hours at which time they were harvested for protein and mRNA analysis. These initial cell experiments were replicated with additional cell lines (SCC25, CAL27, FaDU) which were obtained from ATCC.
3
Comprehensive Reagents and Materials Protocol
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Chloroquine diphosphate (CQ), cycloheximide (CHX), isotretinoin (13-cisRA), NaF, NaHCO3, Na3VO4, Tris-HCl, Triton X-100, Aprotinin, Leupeptin, Pepstatin A, PMSF, Ethanol (molecular biology), Isopropanol (molecular biology), ITS, Puromycin, and 3× FLAG peptide were from Sigma-Aldrich; DTT, EDTA, Formaldehyde (molecular biology), Glycine (molecular biology), IPTG, NaCl, MES, SDS, TAE, Tween-20, FBS, DMEM, IMDM, RPMI-1640, l -Glutamine, Pen Strep, Sodium pyruvate, Trypsin/EDTA, Lipofectamine®, PLUSTM reagent, Proteinase K, RNase A, and Superscript® III First-Strand Synthesis System from ThermoFisher Scientific; Tet-free FBS and Doxycycline from Clontech; Ni-NTA from EMD Millipore; NH2-terminal His-tagged human recombinant OCT4 protein (purity > 90%, made in E. coli) from ProteinONE; COOH-terminal mycDDK-tagged human recombinant MK2 protein from OriGene; MK2 inhibitor PF3644022 from Tocris; MK2 inhibitor III from Santa Cruz Biotechnoloyg; bortezomib from LC Laboratories; p38 inhibitor SB203580 from Sellectchem; Age1-HF, BamH1-HF, EcoR1-HF, Mlu1-HF, Not1-HF, Pme1, Sgf1, Xba1, and Xho1 restriction enzymes from New England Biolabs; bovine serum albumin from Jackson ImmunoResearch Laboratories; All oligonucleotides were synthesized from Integrated DNA Technologies (IDT).
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