Atmi ku55933

Manufactured by Bio-Techne

Sourced in United Kingdom

ATMi KU55933 is a potent and selective inhibitor of the Ataxia Telangiectasia Mutated (ATM) kinase, a key regulator of the cellular response to DNA double-strand breaks. The product is intended for laboratory research use only.

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KU55933 (53 citations)

3 protocols using atmi ku55933

Cell Transfection and Small Molecule Treatments

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JetPEI (PolyPlus), Fugene6 (Promega), Lipofectamine 2000 (Invitrogen), INTERFERin (PolyPlus), Neocarzinostatin (NCS, Sigma) Phleomycin (Sigma), XAV-939 (Tocris), Mimosine (Sigma), RO-3306 (Millipore), ATMi KU55933 (Tocris).

Nagy Z., Kalousi A., Furst A., Koch M., Fischer B, & Soutoglou E. (2016). Tankyrases Promote Homologous Recombination and Check Point Activation in Response to DSBs. PLoS Genetics, 12(2), e1005791.

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ATM-Dependent Co-immunoprecipitation Analyses

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Co-immunoprecipitation analyses were performed as for I-DIRT pull-down with the only exception that protein elution from magnetic beads was performed by incubation with NuPage LDS Sample buffer (Thermo Fisher) supplemented with 50mM DTT for 10 min at 70C. Where indicated, 10 μM ATMi KU55933 (Tocris Bioscience) was added 1h before irradiation. Western blot analysis of protein levels was performed on whole cell lysates prepared by lysis in RIPA buffer supplemented with Complete EDTA free proteinase inhibitor (Roche). The antibodies used for WB analysis are: anti-Rif1 (Di Virgilio et al., 2013 (link)), anti-Flag M2 (Sigma-Aldrich), anti-ZMYND8 (Sigma-Aldrich), anti-γH2AX (Millipore), anti-β Actin (Sigma-Aldrich), and anti-53BP1 (Bethyl).

Delgado-Benito V., Rosen D.B., Wang Q., Gazumyan A., Pai J.A., Oliveira T.Y., Sundaravinayagam D., Zhang W., Andreani M., Keller L., Kieffer-Kwon K.R., Pękowska A., Jung S., Driesner M., Subbotin R.I., Casellas R., Chait B.T., Nussenzweig M.C, & Di Virgilio M. (2018). The Chromatin Reader ZMYND8 Regulates Igh Enhancers to Promote Immunoglobulin Class Switch Recombination. Molecular Cell, 72(4), 636-649.e8.

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Fibroblast Culture and Senescence Induction

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Human dermal fibroblasts used have been described previously (Pekovic et al., 2011 (link)). Human dermal fibroblasts, U2OS and 293T cells were cultured in DMEM containing 10% FBS (Sigma, Gillingham, UK). Human dermal fibroblasts from a patient with lethal foetal akinesis harbouring a homozygous Y259X mutation in the LMNA gene were obtained as an autopsy sample after an informed consent and were a kind gift from Prof. Jos Broers. Conditions to induce quiescence and stress-induced premature senescence have been described previously (Pekovic et al., 2011 (link)). U2OS cells stably expressing GFP-lamin A were selected in G418 (400 μg mL⁻¹). MG-132 (Calbiochem, Merck Millipore, Watford, UK) was used at a concentration of 30 μm and added to culture medium for 6 h. Caffeine (Sigma, Gillingham, UK) was added to cells at a concentration of 10 mm for 1 h prior to irradiation. The ATMi KU-55933 (Tocris Bioscience, Abingdon, UK) was used at a final concentration of 10 μm.

Gibbs-Seymour I., Markiewicz E., Bekker-Jensen S., Mailand N, & Hutchison C.J. (2015). Lamin A/C-dependent interaction with 53BP1 promotes cellular responses to DNA damage. Aging Cell, 14(2), 162-169.

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