Journal: Molecular Cell

Article Title: MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis

doi: 10.1016/j.molcel.2019.02.014

Figure Lengend Snippet:

Article Snippet: DNA-PK inhibitor NU7741 , Cayman Chemical , 14881.

Techniques: Recombinant, Protease Inhibitor, Plasmid Preparation, Staining, Mutagenesis, Mass Spectrometry, Software, Sequencing

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Inhibitors of phosphoinositide 3-kinase (PI3K) and phosphoinositide 3-kinase-related protein kinase family (PIKK)

doi: 10.1080/14756366.2023.2237209

Figure Lengend Snippet: A summary of major PI3K inhibitors.

Article Snippet: DNA-PK inhibitors , Peposertib(M3814) , Merck , DNA-PK: < 3 , I , NCT03770689.

Techniques:

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Inhibitors of phosphoinositide 3-kinase (PI3K) and phosphoinositide 3-kinase-related protein kinase family (PIKK)

doi: 10.1080/14756366.2023.2237209

Figure Lengend Snippet: A summary of major PIKK inhibitors.

Article Snippet: DNA-PK inhibitors , Peposertib(M3814) , Merck , DNA-PK: < 3 , I , NCT03770689.

Techniques:

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Inhibitors of phosphoinositide 3-kinase (PI3K) and phosphoinositide 3-kinase-related protein kinase family (PIKK)

doi: 10.1080/14756366.2023.2237209

Figure Lengend Snippet: Structures of PIKK family members ATR, ATM, and DNA-PKcs inhibitors.

Article Snippet: DNA-PK inhibitors , Peposertib(M3814) , Merck , DNA-PK: < 3 , I , NCT03770689.

Techniques:

Journal: Redox Biology

Article Title: SSBP1 drives high fructose-induced glomerular podocyte ferroptosis via activating DNA-PK/p53 pathway

doi: 10.1016/j.redox.2022.102303

Figure Lengend Snippet: SSBP1 activates DNA-PK to phosphorylate p53 in high fructose-induced glomerular podocytes ferroptosis. (A) Lysates from high fructose-cultured podocytes were incubated with anti -SSBP1, and the co-eluted proteins were examined using DNA-PKcs antibody (n = 3 per group). (B–C) Podocytes were transfected with SSBP1 plasmid and negative control. DNA-PK activity in podocytes was measured by assay kit (n = 6 per group). Western blot analysis of the protein level of DNA-PKcs S2056 phosphorylation in podocytes (n = 6 per group). (D–E) Podocytes were transfected with SSBP1 siRNA and negative control. DNA-PK activity in podocytes was measured by assay kit (n = 6 per group). Western blot analysis of the protein level of DNA-PKcs S2056 phosphorylation in podocytes (n = 6 per group). (F) Western blot analysis of the protein level of DNA-PKcs S2056 phosphorylation in high fructose-stimulated rat glomeruli (n = 6 per group). (G) DNA-PK activity in high fructose-cultured podocytes was measured by assay kit (n = 6 per group). (H) Western blot analysis of the protein level of DNA-PKcs S2056 phosphorylation in high fructose-cultured podocytes (n = 6 per group). (I–J) Podocytes transfected with SSBP1 siRNA or negative control and then cultured with or without fructose (5 mM). DNA-PK activity was measured by assay kit in podocytes (n = 6 per group). Western blot analysis of the protein level of DNA-PKcs S2056 phosphorylation in podocytes (n = 6 per group). (K) Flag-SSBP1, Myc-p53 and HA-DNA-PKcs were expressed alone or in combination in podocytes and Co-IP assays were performed using anti -HA antibody (n = 3 per group). (L–Q) Podocytes were cultured with or without fructose (5 mM) in the presence or absence of vehicle or DNA-PK inhibitor LTURM34. Western blot analysis of the protein levels of p53 S15 phosphorylation, nucleus p53 and SLC7A11 in podocytes (n = 6 per group). Lipid peroxidation in podocytes was analyzed by C11-BODIPY (581/591) staining and measured by flow cytometer (n = 6 per group). The MDA level was measured by assay kit in podocytes (n = 6 per group). Podocyte death was analyzed by propidium iodide staining and measured by flow cytometer (n = 6 per group). Data are plotted as mean ± SEM. P -values were acquired by one-way ANOVA, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: DNA-PK inhibitor LTURM34 was purchased from Glpbio (Montclair, USA).

Techniques: Cell Culture, Incubation, Transfection, Plasmid Preparation, Negative Control, Activity Assay, Measured Assay, Western Blot, Phospho-proteomics, Co-Immunoprecipitation Assay, Staining, Flow Cytometry

Journal: Biochemical and biophysical research communications

Article Title: Inhibiting DNA-PK CS Radiosensitizes Human Osteosarcoma Cells

doi: 10.1016/j.bbrc.2017.03.033

Figure Lengend Snippet: A) RNASeq analysis of DNA-PKCS mRNA in primary bone tumor specimens and cell lines. Specimens included four chondrosarcomas, eight chondroblastomas, five chordoma, five Ewing’s sarcoma (one tissue and four cell lines) and four OS (one tissue and three cell lines). B) Total DNA-PKCS protein levels in OS cell lines compared with HOB cells. C) Levels of DNA-PKCS autophosphorylation at Ser2056 induced with IR (10Gy) and with graded concentration of KU60648, in 143B cells. Results are representative of three independent experiments.

Article Snippet: The DNA-PK CS inhibitor, KU60648, was kindly provided by KuDOS Pharmaceuticals and was dissolved in DMSO as a 1 mM stock solution.

Techniques: Concentration Assay

Journal: Biochemical and biophysical research communications

Article Title: Inhibiting DNA-PK CS Radiosensitizes Human Osteosarcoma Cells

doi: 10.1016/j.bbrc.2017.03.033

Figure Lengend Snippet: Clonogenic survival for 143B cells (A) and U2OS cells (B) treated with vehicle control or 300 nM KU60648 for one hour pre-IR and 24 hours post-IR, rinsed with PBS, media changed and incubated for colony formation. Results are the mean ± SD of three independent experiments fitted to the LQ model.

Article Snippet: The DNA-PK CS inhibitor, KU60648, was kindly provided by KuDOS Pharmaceuticals and was dissolved in DMSO as a 1 mM stock solution.

Techniques: Control, Incubation

Journal: Biochemical and biophysical research communications

Article Title: Inhibiting DNA-PK CS Radiosensitizes Human Osteosarcoma Cells

doi: 10.1016/j.bbrc.2017.03.033

Figure Lengend Snippet: A–C) FACS histograms for U2OS cells treated with vehicle control (A), 5 Gy (B), and 5 Gy plus 100 nM KU60648 (C). Results are representative of three independent experiments. D–F) The summary of the cell cycle analyses for 143B cells (D), U2OS cells (E) and HOB cells (F). Results are mean ± SD of three or more independent experiments. (* P < 0.05)

Article Snippet: The DNA-PK CS inhibitor, KU60648, was kindly provided by KuDOS Pharmaceuticals and was dissolved in DMSO as a 1 mM stock solution.

Techniques: Control

Journal: Biochemical and biophysical research communications

Article Title: Inhibiting DNA-PK CS Radiosensitizes Human Osteosarcoma Cells

doi: 10.1016/j.bbrc.2017.03.033

Figure Lengend Snippet: Representative confocal microscopy images of γH2AX foci 24 hours post-IR in cells treated with vehicle control, 5 Gy or 5 Gy plus 100 nM KU60648 (added 1 hour before IR) in 143B cells (A) and U2OS cells (B). Quantification of percentage of cells with > 20 γH2AX foci in cells treated with vehicle control, 100 nM KU60648 alone, 5 Gy or 5 Gy plus 100 nM KU60648 in 143B cells (C) and U2OS cells (D). Results are mean ± SD from 2 independent experiments. For each treatment group, >150 and >90 nuclei were counted for 143B and U2OS cells, respectively.

Article Snippet: The DNA-PK CS inhibitor, KU60648, was kindly provided by KuDOS Pharmaceuticals and was dissolved in DMSO as a 1 mM stock solution.

Techniques: Confocal Microscopy, Control