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Sb218078

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SB218078 is a chemical compound that inhibits the activity of the Cdc7 protein kinase. It is commonly used in biological research as a tool to study the role of Cdc7 in cellular processes.

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Lab products found in correlation

Nu7441, by Bio-Techne (2 mentions) Nocodazole, by Merck Group (1 mentions) Mk 1775, by Selleck Chemicals (1 mentions) Rpa70, by Abcam (1 mentions) Roscovitine, by Selleck Chemicals (1 mentions) α tubulin, by Merck Group (1 mentions) Chk2 pt68, by Cell Signaling Technology (1 mentions) Ve 821, by Selleck Chemicals (1 mentions) Rpa32, by Fortis Life Sciences (1 mentions) Rpa32 pt21, by Abcam (1 mentions) Rpa32 ps4 8, by Fortis Life Sciences (1 mentions) Chk1 ps345, by Cell Signaling Technology (1 mentions) Thymidine, by Merck Group (1 mentions) Z vad fmk, by Promega (1 mentions) Pf477736, by Bio-Techne (1 mentions) Nutlin 3, by Bio-Techne (1 mentions) Ci 1040, by Selleck Chemicals (1 mentions) Danusertib, by Selleck Chemicals (1 mentions) Ku55933, by Selleck Chemicals (1 mentions) Plx4720, by Selleck Chemicals (1 mentions)

6 protocols using sb218078

1

Characterizing CHK1-Mediated Histone Acetylation

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Full length p300 was expressed in Sf9 cells and purified with 3×FLAG beads. Both recombinant human CHK1 protein and recombinant mononucleosomes were purchased from Active Motif (31163, 81070). First, H3.3 mononucleosomes were incubated with CHK1(50 ng) in kinase buffer (50 mM Tris-HCl pH 7.4, 10 mM MgCl2, 1 mM DTT, 5% glycerol, protease inhibitor cocktail) in the presence or absence of 0.2 mM ATP at 37 °C for 5 min. The reaction was terminated by the addition of 5 μM CHK1 inhibitor SB218078 (Tocris, Cat. No. 2560). Immediately, the HAT assay was performed by adding 25 uM acetyl-CoA (3H AcetylCOA in the case of HOT reaction), 100 ng of full length p300 (plus H3.1 mononucleosomes in case of the trans assay) for 30 min at 30 °C. The reaction was stopped by adding SDS sample buffer and the samples were subject to downstream analysis (immunoblot analysis, autoradiography, mass spectrometry).
Martire S., Gogate A.A., Whitmill A., Tafessu A., Nguyen J., Teng Y.C., Tastemel M, & Banaszynski L.A. (2019). Phosphorylation of histone H3.3 at serine 31 promotes p300 activity and enhancer acetylation. Nature genetics, 51(6), 941-946.
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2

Characterizing CHK1-Mediated Histone Acetylation

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Full length p300 was expressed in Sf9 cells and purified with 3×FLAG beads. Both recombinant human CHK1 protein and recombinant mononucleosomes were purchased from Active Motif (31163, 81070). First, H3.3 mononucleosomes were incubated with CHK1(50 ng) in kinase buffer (50 mM Tris-HCl pH 7.4, 10 mM MgCl2, 1 mM DTT, 5% glycerol, protease inhibitor cocktail) in the presence or absence of 0.2 mM ATP at 37 °C for 5 min. The reaction was terminated by the addition of 5 μM CHK1 inhibitor SB218078 (Tocris, Cat. No. 2560). Immediately, the HAT assay was performed by adding 25 uM acetyl-CoA (3H AcetylCOA in the case of HOT reaction), 100 ng of full length p300 (plus H3.1 mononucleosomes in case of the trans assay) for 30 min at 30 °C. The reaction was stopped by adding SDS sample buffer and the samples were subject to downstream analysis (immunoblot analysis, autoradiography, mass spectrometry).
Martire S., Gogate A.A., Whitmill A., Tafessu A., Nguyen J., Teng Y.C., Tastemel M, & Banaszynski L.A. (2019). Phosphorylation of histone H3.3 at serine 31 promotes p300 activity and enhancer acetylation. Nature genetics, 51(6), 941-946.
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3

Investigating Cell Cycle Regulation

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The following chemical inhibitors were used at the indicated concentrations unless stated otherwise: WEE1 inhibitor MK-1775 (500 nM; Selleck Chemicals), CHK1 inhibitor SB 218078 (2 μM; Tocris), ATR inhibitor VE-821 (20 μM; Selleck Chemicals), CDK1 inhibitor RO-3306 (10 μM; Calbiochem), CDK inhibitor Roscovitine (20 μM; Selleck Chemicals), thymidine (2 mmol/L; Sigma-Aldrich), and nocodazole (100 ng/mL; Sigma-Aldrich), IdU (10 μg/ml, Sigma-Aldrich), CIdU (10 μg/ml, Sigma-Aldrich). The following antibodies were used: CHK1 (Santa Cruz Biotechnology), CHK1-pS345 (Cell Signaling), CHK2 (Santa Cruz Biotechnology), CHK2-pT68 (Cell Signaling), phospho-Histone-H2AX-Ser139 (γH2AX; Millpore), CDK1 (Santa Cruz Biotechnology), CDK1-pTyr15 (Cell Signaling), RPA70 (Abcam), phospho-(Ser) CDKs Substrate (pCDK Substrate; Cell Signaling), CDK2 (Santa Cruz Biotechnology), CDK2-pTyr15 (Abcam), RPA32-pT21 (Abcam), RPA32-pS33 (Abcam), RPA32-pS4/8 (Bethyl Laboratories), RPA32 (Bethyl Laboratories), WEE1 (Santa Cruz Biotechnology), α-Tubulin (Sigma-Aldrich), CIdU (rat anti-BrdU; Accurate Chemical), IdU (mouse anti-BrdU; Becton Dickinson).
Zheng H., Shao F., Martin S., Xu X, & Deng C.X. (2017). WEE1 inhibition targets cell cycle checkpoints for triple negative breast cancers to overcome cisplatin resistance. Scientific Reports, 7, 43517.
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4

Small Molecule Inhibitors in Cell Culture

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2 mM HU (Millipore Sigma, Cat#H8627) was prepared in culture medium
prior to each experiment. Nutlin-3 (Tocris, Cat#3984), Pifithrin-α (Focus
Biomolecules, Cat#10–2480), SB 218078 (Tocris, Cat#2560), PF477736
(Tocris, Cat#4277) and zVAD-FMK (Promega, Cat# G7231) stocks were prepared in
DMSO and kept at −20ºC until needed, when dilutions were prepared
in culture medium for cell treatment for final concentrations of 10 μM,
10 μM, 30 nM, 5nM and 50 μM, respectively.
Dwivedi V.K., Pardo-Pastor C., Droste R., Kong J.N., Tucker N., Denning D.P., Rosenblatt J, & Horvitz H.R. (2021). Replication stress promotes cell elimination by extrusion. Nature, 593(7860), 591-596.
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5

DNA Damage Repair Inhibitors

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The DNA-PK inhibitor (NU7441, Tocris Bioscience, UK), ATM inhibitor (KU55933, Calbiochem, Germany), and Chk1/2 inhibitor (SB218078, Tocris Bioscience, UK) were used to treat cells at final concentrations of 10, 10, and 2.5 μM, respectively. The efficacy of these inhibitors, including in the inhibition of cell cycle checkpoints and/or DSB repair, is well established19 (link)51 (link)52 (link).
Yamauchi M., Shibata A., Suzuki K., Suzuki M., Niimi A., Kondo H., Miura M., Hirakawa M., Tsujita K., Yamashita S, & Matsuda N. (2017). Regulation of pairing between broken DNA-containing chromatin regions by Ku80, DNA-PKcs, ATM, and 53BP1. Scientific Reports, 7, 41812.
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6

Screening of Protein Kinase Inhibitors for TNBC

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The fifty-five protein kinase inhibitors (PKIs) were purchased from following sources: BML-275, FR 180204, IKK16, GW 843682X, NSC 109555, NU7441, PD407824, PF 573228, SB 218078, TCS PIM-1-1, TCS PIM-1-4a, and TPCA-1 from Tocris Biosciences (Bristol, UK); indirubin-3′-monoxime and Ro-31-8220 from Calbiochem (San Diego, CA, USA); A-769662, bosutinib, chelerythrine, CP690550, fasudil, gefitinib, imatinib, nilotinib, PKC412, roscovitine, SNS-314, and tozasertib from LC Laboratories (Woburn, MA, USA); AT7867, AT9283, AZD1152, AZD1480, BI 2536, BIX 02189, CHIR-99021, CI-1040, CYC116, danusertib, enzastaurin, GDC-0879, INCB018424, JNJ-7706621, KU-55933, LY2228820, MLN8237, PD-0325901, PF-4708671, PLX-4032, PLX-4720, SB216763, SNS-032, SP600125, VX-702, Y-27632, and ZM447439 from Selleck Chemicals (Houston, TX, USA); U0126 from Promega (Madison, WI, USA); TBCA from Millipore (Burlington, MA, USA).
All TNBC cells in this study were obtained from the American Type Culture Collection (Manassas, VA, USA). The cultured cells were monitor by trypan blue cell counting as described previously [58 (link)].
Yi Y.W., You K.S., Han S., Ha I.J., Park J.S., Lee S.G, & Seong Y.S. (2022). Inhibition of IκB Kinase Is a Potential Therapeutic Strategy to Circumvent Resistance to Epidermal Growth Factor Receptor Inhibition in Triple-Negative Breast Cancer Cells. Cancers, 14(21), 5215.
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