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Gsk2830371

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GSK2830371 is a laboratory compound developed by MedChemExpress. It is a chemical tool used for research purposes. The core function of GSK2830371 is to inhibit the activity of the protein PARP14. Further details on the intended use or application of this compound are not available.

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Lab products found in correlation

Ku 55933, by MedChemExpress (2 mentions) Ppm1d, by Santa Cruz Biotechnology (2 mentions) Phospho histone h2a x ser139 clone d7t2v, by Cell Signaling Technology (2 mentions) Kap1 s824 4127 and ppm1d clone d4f7, by Cell Signaling Technology (2 mentions) Flag tag (flag), by Roche (2 mentions) Ve 821, by MedChemExpress (2 mentions) Nu7026, by MedChemExpress (2 mentions) γh2ax, by Roche (2 mentions) Camptothecin, by MedChemExpress (1 mentions) γh2ax 05 636, by Merck Group (1 mentions) Alexa fluor conjugated antibody, by Thermo Fisher Scientific (1 mentions) Mitomycin c, by Santa Cruz Biotechnology (1 mentions) Olaparib, by MedChemExpress (1 mentions) Resazurin, by Merck Group (1 mentions) Neocarzinostatin ncs, by Merck Group (1 mentions) Carboxyfluorescein diacetate succinimidyl ester cfse, by Merck Group (1 mentions) Fitc conjugated anti brdu antibody, by BD (1 mentions) γh2ax, by Cell Signaling Technology (1 mentions) Nutlin 3, by MedChemExpress (1 mentions) P ser15 p53, by Santa Cruz Biotechnology (1 mentions)

5 protocols using gsk2830371

1

Antibody Detection and Validation Protocol

Cited 1 time
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The following antibodies were used in this study: TRF2 (ab108997, for WB), TIN2 (ab197894, for WB) from Abcam; TRF2 (NB110-57130, for IF), TIN2 (NBP2-55709, for IF), RAP1 (NBP1-82433, for IF), 53BP1 (NB100-305, for IF) from Novus Biologicals; TRF2 (sc271710, for IF), TIN2 (sc73177, for IF), TPP1 (sc100597, for IF and WB), RAP1 (sc53434, for WB), PPM1D (sc376257, for IF and WB), PPM1D (sc20712, for IF) from Santa Cruz Biotechnology; Phospho-Histone H2A.X (Ser139) (clone D7T2V, #80312), KAP1-S824 (#4127) and PPM1D (clone D4F7, 11901 for WB) from Cell Signaling Technology; γH2AX (05-636, for WB), GFP (11814460001, for WB), FLAG (F1804, for IF), Fk2 (04-263, for IF) from Roche. A custom-made pTRF2-S410 antibody was generated by immunization of rabbits with KLH-conjugated phospho-peptide RLVLEEDpSQSTEPSA corresponding to amino acids 403–417 of the human TRF2 (according to the numbering in reference sequence NP_005643.2) (Davids Biotechnologie). Subsequently, immune sera was affinity purified using negative and positive selection with non-phosphorylated and phosphorylated peptides, respectively. PPM1D inhibitor GSK2830371 was from MedChemExpress and was validated previously (44 (link),49 (link)). Validated small molecule inhibitors of ATM (KU-55933), ATR (VE-821) and DNA-PK (NU7026) were from MedChemExpress and were used at final concentrations 10, 10 and 5 μM, respectively.
Storchova R., Palek M., Palkova N., Veverka P., Brom T., Hofr C, & Macurek L. (2023). Phosphorylation of TRF2 promotes its interaction with TIN2 and regulates DNA damage response at telomeres. Nucleic Acids Research, 51(3), 1154-1172.
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2

DNA Damage Response Pathway Profiling

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Following antibodies were used: WIP1 antibody (clone F-10, sc-376257), p21 (sc-397), p53 (clone D01, sc-126), BRCA1 (sc-6954), rabbit-53BP1 (sc-22760), RAD51 (sc-6862) and TFIIH (sc-293, used as loading control) from Santa Cruz Biotechnology (Dallas, TX, USA); phoshpo-Thr543-53BP1 (#3428), phospho-S15-p53 (#9284) from Cell Signaling Technology (Danvers, MA, USA); RPA2 (clone 9H8, ab2175), and phospho-Ser1524-BRCA1 (ab2401) from Abcam (Cambridge, UK), γH2AX (05-636), and mouse monoclonal 53BP1 (MAB3802) from Merck Millipore (Burlington, MA, USA); phospho-S824-KAP1 (GTX63711), KAP1 (GTX62973) and PP4C (GTX114659) from Genetex (Irvine, CA, USA); secondary Alexa Fluor conjugated antibodies from Thermo Fisher Scientific (Waltham, MA, USA). WIP1 inhibitor GSK2830371 (here referred to as WIP1i and used at 0.5 μM unless stated otherwise), olaparib and camptothecin (all Medchemexpress, New York, NJ, USA) and mitomycin C (Santa Cruz Biotechnology, Dallas, TX, USA) were dissolved in DMSO and used at indicated concentrations.
Burdova K., Storchova R., Palek M, & Macurek L. (2019). WIP1 Promotes Homologous Recombination and Modulates Sensitivity to PARP Inhibitors. Cells, 8(10), 1258.
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3

Antibody Detection and Validation Protocol

Cited 1 time
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The following antibodies were used in this study: TRF2 (ab108997, for WB), TIN2 (ab197894, for WB) from Abcam; TRF2 (NB110-57130, for IF), TIN2 (NBP2-55709, for IF), RAP1 (NBP1-82433, for IF), 53BP1 (NB100-305, for IF) from Novus Biologicals; TRF2 (sc271710, for IF), TIN2 (sc73177, for IF), TPP1 (sc100597, for IF and WB), RAP1 (sc53434, for WB), PPM1D (sc376257, for IF and WB), PPM1D (sc20712, for IF) from Santa Cruz Biotechnology; Phospho-Histone H2A.X (Ser139) (clone D7T2V, #80312), KAP1-S824 (#4127) and PPM1D (clone D4F7, 11901 for WB) from Cell Signaling Technology; γH2AX (05-636, for WB), GFP (11814460001, for WB), FLAG (F1804, for IF), Fk2 (04-263, for IF) from Roche. A custom-made pTRF2-S410 antibody was generated by immunization of rabbits with KLH-conjugated phospho-peptide RLVLEEDpSQSTEPSA corresponding to amino acids 403–417 of the human TRF2 (according to the numbering in reference sequence NP_005643.2) (Davids Biotechnologie). Subsequently, immune sera was affinity purified using negative and positive selection with non-phosphorylated and phosphorylated peptides, respectively. PPM1D inhibitor GSK2830371 was from MedChemExpress and was validated previously (44 (link),49 (link)). Validated small molecule inhibitors of ATM (KU-55933), ATR (VE-821) and DNA-PK (NU7026) were from MedChemExpress and were used at final concentrations 10, 10 and 5 μM, respectively.
Storchova R., Palek M., Palkova N., Veverka P., Brom T., Hofr C, & Macurek L. (2023). Phosphorylation of TRF2 promotes its interaction with TIN2 and regulates DNA damage response at telomeres. Nucleic Acids Research, 51(3), 1154-1172.
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4

Evaluating DNA Damage and Cell Cycle Responses

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The following antibodies were used: WIP1 (sc-130655), p53 (sc-6243), TFIIH (sc-293), importin (sc-137016), p21 (sc-397) from Santa Cruz; pSer15-p53 (#9284), γH2AX (#9718), p38 MAPK Thr180/Tyr182 (#9216S) and p38 MAPK (#9212) from Cell Signaling Technology); γH2AX (05-636, Millipore); MDM2 (Calbiochem); Alexa Fluor-labelled secondary antibodies (Life Technologies); anti-BrdU FITC-conjugated antibody (#347583, BD Biosciences) and anti-pSer10-H3 antibody (Upstate). Doxorubicin hydrochloride (Sigma), GSK2830371 and nutlin-3 (both MedChem Express) were diluted in DMSO and used at indicated doses. Resazurin, neocarzinostatin (NCS) and carboxyfluorescein diacetate succinimidyl ester (CFSE) were purchased from Sigma.
Pechackova S., Burdova K., Benada J., Kleiblova P., Jenikova G, & Macurek L. (2016). Inhibition of WIP1 phosphatase sensitizes breast cancer cells to genotoxic stress and to MDM2 antagonist nutlin-3. Oncotarget, 7(12), 14458-14475.
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5

PPM1D Inhibition Enhances Radiation Sensitivity

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PPM1D-mutant DMG cell lines BT869 and SF776 were treated with vehicle control (DMSO), 10 μM of PPM1D inhibitor GSK2830371 (MedChemExpress, 5140), 2 Gy ionizing radiation treatment, or the combination of both. Cells were seeded at a density of 1000 cells per well in 96-well ULA plates. Each biological replicate had six technical replicates for each cell line. The proliferation rates were analyzed over time using spheroid assay in the Incucyte live cell imaging system. An unpaired t-test assuming equal variance was conducted to compare the fold change values between different conditions at day 5 of the experiment.
Khadka P., Reitman Z.J., Lu S., Buchan G., Gionet G., Dubois F., Carvalho D.M., Shih J., Zhang S., Greenwald N.F., Zack T., Shapira O., Pelton K., Hartley R., Bear H., Georgis Y., Jarmale S., Melanson R., Bonanno K., Schoolcraft K., Miller P.G., Condurat A.L., Gonzalez E.M., Qian K., Morin E., Langhnoja J., Lupien L.E., Rendo V., Digiacomo J., Wang D., Zhou K., Kumbhani R., Guerra Garcia M.E., Sinai C.E., Becker S., Schneider R., Vogelzang J., Krug K., Goodale A., Abid T., Kalani Z., Piccioni F., Beroukhim R., Persky N.S., Root D.E., Carcaboso A.M., Ebert B.L., Fuller C., Babur O., Kieran M.W., Jones C., Keshishian H., Ligon K.L., Carr S.A., Phoenix T.N, & Bandopadhayay P. (2022). PPM1D mutations are oncogenic drivers of de novo diffuse midline glioma formation. Nature Communications, 13, 604.
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