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Ku 55933

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KU-55933 is a selective inhibitor of the serine/threonine protein kinase ATM (Ataxia Telangiectasia Mutated). It functions by blocking the enzymatic activity of ATM, which is involved in the cellular response to DNA double-strand breaks.

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Lab products found in correlation

Sb203580, by Selleck Chemicals (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Hydroethidine, by Thermo Fisher Scientific (1 mentions) Anti phospho histone h2a x ser139, by Merck Group (1 mentions) Anti nrf2, by Abcam (1 mentions) Anti phospho atm ser1981, by Merck Group (1 mentions) Anti heme oxygenase 1 ho 1, by Proteintech (1 mentions) Anti phospho erk1 2, by Cell Signaling Technology (1 mentions) Tert butylhydroquinone tbhq, by Merck Group (1 mentions) U0126, by Promega (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Nucleofector 2 device, by Lonza (1 mentions) Amaxa nucleofector kit, by Lonza (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Rpmi medium, by Thermo Fisher Scientific (1 mentions) Polyethylenimine (pei), by Polysciences (1 mentions) Nutlin 3, by Selleck Chemicals (1 mentions) Trametinib, by Santa Cruz Biotechnology (1 mentions) Gefitinib, by Santa Cruz Biotechnology (1 mentions)

7 protocols using ku 55933

1

Immunohistochemical Analysis of NRF2 Signaling

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Antibodies used were anti-NRF2 (Abcam Cambridge, UK), anti-phospho-histone H2A.X (Ser139), anti-phospho-ATM (Ser1981) (Merck KGaA, Darmstadt, Germany), anti phospho-ERK 1/2 (Cell Signaling Technology, Danvers, MA, USA), and anti-heme oxygenase 1 (HO-1) (Proteintech, Rosemont, IL, USA). ThermoFisher Scientific provided the Alexa Fluor-488 conjugated Goat Anti Rabbit and Alexa Fluor-555 conjugated Goat Anti Mouse secondary antibodies. The MEK 1/2 chemical inhibitor U0126 was purchased from Promega (Beijing, China). The ATM chemical inhibitor ku55933 was from Santa Cruz (Dallas, TX, USA). The chemical inducer of NRF2 tert-butylhydroquinone (tBHQ) was purchased from Sigma (Merck KGaA, Darmstadt, Germany). Hoechst 33342 and hydroethidine were purchased from ThermoFisher Scientific.
Wang J., Oikawa M, & Konishi T. (2023). Stimulation of Nuclear Factor (Erythroid-Derived 2)-like 2 Signaling by Nucleus Targeted Irradiation with Proton Microbeam. Biology, 12(3), 419.
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2

Cell Line Characterization and Transfection

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293T (human embryonic kidney), DLD-1, HCT-116 (colorectal carcinoma), MCF7, and BT474 (breast cancer) cells were obtained from ATCC; PC9 cells (NSCLC) were obtained from ECACC-Sigma-Aldrich; and Kelly cells (neuroblastoma) were a kind gift from Dr. C. Einvik. HCT-116 and MCF7 cells are WT for TP53 (http://p53.iarc.fr/CellLines.aspx). All cells were grown in DMEM (Life Technologies), except Kelly and PC9 cells, grown in RPMI medium (Life Technologies), both supplemented with 10% fetal bovine serum (Life Technologies) and 0.6% penicillin/streptomycin (Life Technologies). Cells were transfected with a Nucleofector II device (Lonza) using the Amaxa Nucleofector kit (Lonza) and electroporation program recommended by the manufacturer. 293T cells were transfected using polyethylenimine (Polysciences). The efficiency of each transfection was assessed in parallel using a GFP-containing plasmid.
Nutlin 3 was purchased from SelleckChem, and doxorubicin was purchased from Abcam. Gefitinib, WZ4002, TAE684, trametinib, and KU-55933 were purchased from Santa Cruz Biotechnology.
Guernet A., Mungamuri S.K., Cartier D., Sachidanandam R., Jayaprakash A., Adriouch S., Vezain M., Charbonnier F., Rohkin G., Coutant S., Yao S., Ainani H., Alexandre D., Tournier I., Boyer O., Aaronson S.A., Anouar Y, & Grumolato L. (2016). CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and Functional Analysis of Oncogenic Driver Mutations. Molecular cell, 63(3), 526-538.
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3

Chemical Compounds for Experimental Research

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KU55933 was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA), SB203580 was purchased from Selleck Chemicals (Houston, TX, USA), NS-398 was purchased from Abcam (Cambridge, UK) and protoporphyrin IX zinc (II) (ZnPP) was purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). These chemicals were dissolved in 100% dimethyl sulfoxide and stored in small aliquots at 20°C. Protease inhibitor cocktail and protein inhibitor cocktail were purchased from Roche Diagnostics GmbH (Mannheim, Germany) and Sigma-Aldrich (Merck KGaA), respectively.
Yang X., Liu H., Jiang X., Jin C., Xu Z., Li T., Wang Z, & Wang J. (2018). Cyclooxygenase-2-mediated upregulation of heme oxygenase 1 mitigates the toxicity of deuterium-tritium fusion radiation. International Journal of Molecular Medicine, 42(4), 1945-1954.
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4

Antiviral Agent Inhibition Assay

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Two h after inoculation with EVA71 (MOI = 1), cells were gently rinsed once with PBS to eliminate virions remained in the medium and then treated with different doses of ATR inhibitor (VE-821, A2521; ApexBio, TX, USA), DNA-PK inhibitor (KU-57788, S2638; Selleck, TX, USA), or ATM inhibitor (KU-55933, SC202963; Santa Cruz), respectively.
Yu J., Zhang W., Huo W., Meng X., Zhong T., Su Y., Liu Y., Liu J., Wang Z., Song F., Zhang S., Li Z., Yu X., Yu X, & Hua S. (2022). Regulation of host factor γ-H2AX level and location by enterovirus A71 for viral replication. Virulence, 13(1), 241-257.
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5

Targeted Inhibition of DNA Damage Response

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Deferoxamine, hydroxyurea, and mitomycin C (Sigma) were dissolved in H2O and used at final concentrations of 250 μM, 2 mM, and 1 μM, respectively. N-Ethylmaleimide (Sigma) was dissolved in EtOH and added to lysis buffer at a final concentration of 4 mM. VE-821 (Vertex Pharmaceuticals), KU-55933 (Santa Cruz), and NU-7441 (Tocris Bioscience) were dissolved in DMSO and used at final concentrations of 1 μM, 10 μM, and 1 μM, respectively. Cisplatin (Sigma) was dissolved in DMF and used at a final concentration of 20 μg/mL.
Scanlon S.E, & Glazer P.M. (2014). Hypoxic Stress Facilitates Acute Activation and Chronic Down-Regulation of Fanconi Anemia Proteins. Molecular cancer research : MCR, 12(7), 1016-1028.
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6

Molecular Mechanisms of Cellular Stress Response

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Cycloheximide (CHX) was from Calbiochem (Darmstadt, Germany). SB203580 was from Selleck (Houston, TX, USA). Actinomycin D was from Solarbio (Beijing, China). Ku55933 was from Santa Cruz (Dallas, TX, USA). The following primary antibodies were used: TFAM, β‐actin, proliferating cell nuclear antigen (Santa Cruz Biotechnology), α‐tubulin (Abcam, Cambridge, UK), p38 (pT180/pY182; Becton Dickinson, San Jose, CA, USA), ATM (pSer1981; Calbiochem), HuR (Millipore, Darmstadt, Germany), cleaved caspase 3 (Cell Signaling Technology, Danvers, MA, USA), B‐cell lymphoma 2, and BAX (Proteintech, Wuhan, China). The siRNA oligonucleotides were synthesized by GenePharma (Shanghai, China) and DNA primers were synthesized by General Biosystems (Chuzhou, China) (Table 1). TFAM shRNAs were purchased from OriGene (Rockville, MD, USA).
Zhang R, & Wang J. (2018). HuR stabilizes TFAM mRNA in an ATM/p38‐dependent manner in ionizing irradiated cancer cells. Cancer Science, 109(8), 2446-2457.
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7

Molecular regulation of HIC1 and MTA1

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The expression vectors for full-length FLAG-HIC1, the non-SUMOylatable FLAG-HIC1 E316A, for His-SUMO2, Myc-MTA1, the non SUMOylatable Myc-MTA1 K509R and SIM-deficient Myc-MTA1 AAA mutants have been described previously [8 (link), 10 (link), 19 (link)]. The ΔMKHEP deletion mutant was generated by the two-round PCR mutagenesis strategy.
Etoposide and the Chk2 inhibitor C3742 (Chk2 inhibitor II hydrate) were purchased from Sigma-Aldrich. Wortmannin (Calbiochem), a PI3K inhibitor which also inhibits PIKKs and ATM (KU-55933; Santa Cruz Biotechnology) and DNA-PKcs (NU-7441, Selleckchem) inhibitors were dissolved in DMSO and used at a final concentration of 10 μM. Inhibitors were added to culture medium 1 hour before subsequent treatments.
Paget S., Dubuissez M., Dehennaut V., Nassour J., Harmon B.T., Spruyt N., Loison I., Abbadie C., Rood B.R, & Leprince D. (2016). HIC1 (hypermethylated in cancer 1) SUMOylation is dispensable for DNA repair but is essential for the apoptotic DNA damage response (DDR) to irreparable DNA double-strand breaks (DSBs). Oncotarget, 8(2), 2916-2935.
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