chk1 kinase  (Carna Inc)

Journal: Clinical and Translational Radiation Oncology

Article Title: Overexpression of JAML in colorectal cancer cells predicts higher radiosensitivity by inactivating ATR pathway

doi: 10.1016/j.ctro.2025.101016

Figure Lengend Snippet: CRC cells with JAML overexpression demonstrate increased apoptosis and DNA damage after irradiation, accompanied by inhibiting the ATR-CHK1 signaling pathway. (A) Representative images of apoptosis of LOVO and DLD-1 cells using flow cytometry after 24 h after X-ray irradiation. (B and C) Quantitative statistics of apoptosis of LOVO and DLD-1 cells. (D) Representative images of fluorescence of γH2AX in LOVO and DLD-1 cells after 12 h of X-ray irradiation or lack of X-ray irradiation. (E and F) Quantitative statistical analysis of the γH2AX-positive rate in LOVO and DLD-1 cells. (G and L) Western blot images of the ATR-CHK1 signaling pathway’s protein expression in LOVO and DLD-1 cells after 24 h of X-ray irradiation/lack of irradiation. (H–K and M−P) Quantitative statistical analysis of the relative ATR-CHK1 signaling pathway’s protein expression in LOVO and DLD-1 cells (*, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; and ****, P ≤ 0.0001).

Article Snippet: Antibodies against GAPDH (Proteintech, dilution 1:50000, China), pCHK1 (Cell Signaling Technology, dilution 1:1000, USA), checkpoint kinase 1 (CHK1) (MCE, dilution 1:1000, MCE), pATR (Cell Signaling Technology, dilution 1:1000, USA), and ataxia telangiectasia and Rad3-related protein (ATR) (Proteintech, dilution 1:1000, China) were used.

Techniques: Over Expression, Irradiation, Flow Cytometry, Fluorescence, Western Blot, Protein-Protein interactions, Expressing

Journal: Clinical and Translational Radiation Oncology

Article Title: Overexpression of JAML in colorectal cancer cells predicts higher radiosensitivity by inactivating ATR pathway

doi: 10.1016/j.ctro.2025.101016

Figure Lengend Snippet: CRC tumors with JAML overexpression show lower Ki67 expression after X-ray radiation therapy , accompanied by inhibiting the ATR-CHK1 signaling pathway in vivo. (A)Representative immunofluorescence images of Ki67 in LOVO and DLD-1 tumor tissues. (B and C) Quantitative statistical analysis of the expression of Ki67 in LOVO and DLD-1 tumor tissues. (D) Representative western blot images of the ATR-CHK1 signaling pathway in LOVO and DLD-1 tumor tissues. (E–L) Quantitative statistical analysis of the relative ATR-CHK1 signaling pathway’s protein expression in LOVO and DLD-1 tumor tissues (*, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; and ****, P ≤ 0.0001).

Article Snippet: Antibodies against GAPDH (Proteintech, dilution 1:50000, China), pCHK1 (Cell Signaling Technology, dilution 1:1000, USA), checkpoint kinase 1 (CHK1) (MCE, dilution 1:1000, MCE), pATR (Cell Signaling Technology, dilution 1:1000, USA), and ataxia telangiectasia and Rad3-related protein (ATR) (Proteintech, dilution 1:1000, China) were used.

Techniques: Over Expression, Expressing, In Vivo, Immunofluorescence, Western Blot, Protein-Protein interactions

Journal: bioRxiv

Article Title: Phosphorylation of Claspin by elF2α kinase protects cells from heat stress

doi: 10.1101/2025.08.21.670878

Figure Lengend Snippet: ( A ) Kinase prediction for the C-terminal phosphorylation sites (right, S1325; left, T1326) of Claspin under heat shock stress. ( B ) Wild-type, 4KO (lacking PERK, PKR, HRI, and GCN2), 4KO + PERK(stably expressed), 4KO + PKR, 4KO + HRI, and 4KO + GCN2 MEF cells were cultured at 42°C for 4 hrs. Whole cell extracts were harvested and analyzed by Western blotting with the indicated antibodies. ( C ) The same set of cells as in B were transfected with or without siHRI or siGCN2 for 24 hrs, and then cultured at 42°C for 4 or 12 hrs, and were harvested. Whole cell extracts were analyzed by Western blotting with the indicated antibodies.

Article Snippet: Purified Chk1 (02–117), HRI (05–154), and GCN2 (05–153) kinases were obtained from Carna Bioscience.

Techniques: Phospho-proteomics, Stable Transfection, Cell Culture, Western Blot, Transfection

Journal: bioRxiv

Article Title: Phosphorylation of Claspin by elF2α kinase protects cells from heat stress

doi: 10.1101/2025.08.21.670878

Figure Lengend Snippet: ( A ) Purified wild-type, ST/A, and ST/E mutant Claspin proteins were treated with λ phosphatase (λPPase), pre-treated with PhosSTOP™, and then mixed with purified HRI or GCN2 (20 ng each) and incubated in kinase buffer at 30°C for 30 min. ( B ) Wild-type Chk1 (WT) or a kinase-dead Chk1 mutant (D130A, KD) protein was mixed with HRI or GCN2 kinase (20 ng each) and incubated in kinase buffer with CHKtide polypeptide at 30°C for 60 min. In ( A ) and ( B ), the reaction mixtures were subjected to SDS-PAGE. The gel was stained with Coomassie Brilliant Blue (CBB) (right panel) and then autoradiographed (left panel).

Article Snippet: Purified Chk1 (02–117), HRI (05–154), and GCN2 (05–153) kinases were obtained from Carna Bioscience.

Techniques: Purification, Mutagenesis, Incubation, SDS Page, Staining

Journal: bioRxiv

Article Title: Phosphorylation of Claspin by elF2α kinase protects cells from heat stress

doi: 10.1101/2025.08.21.670878

Figure Lengend Snippet: ( A ) HCT116 Claspin-myc-AID cells stably expressing wild-type Claspin-Flag were treated with auxin and with or without GCN2 inhibitor (GCN2-IN-1, A-92) for 3 hrs. The cells were then cultured at 37°C or 42°C for 90 min. Whole cell extracts were harvested and analyzed by Western blotting using the anti-DDDK antibody. ( B ) HCT116 Claspin-myc-AID cells, treated with or without 0.5 mM auxin and 10 nM GCN2 inhibitor for 3 hrs, were cultured at 37°C or 42°C for 90 min. Cells were plated at a density of 500 cells per dish in medium with or without auxin. After 3 weeks, colonies were stained with Coomassie Brilliant Blue (CBB) (left) and counted (right). Statistical significance is indicated as follows: ns (p > 0.05); * (p ≤ 0.05); ** (p ≤ 0.01); *** (p ≤ 0.001).

Article Snippet: Purified Chk1 (02–117), HRI (05–154), and GCN2 (05–153) kinases were obtained from Carna Bioscience.

Techniques: Stable Transfection, Expressing, Cell Culture, Western Blot, Staining

Journal: bioRxiv

Article Title: Phosphorylation of Claspin by elF2α kinase protects cells from heat stress

doi: 10.1101/2025.08.21.670878

Figure Lengend Snippet: ( A ) HCT116 Claspin-myc-AID cells, stably expressing wild-type (WT), ST/A, or ST/E Claspin, were treated with auxin in the presence and absence of ATR inhibitor (ATRi; AZD6738) for 3 hrs. Cells were then cultured at either 37°C or 42°C for 90 min. Whole-cell extracts were collected and analyzed via western blotting with the indicated antibodies. ( B ) The same cells used in ( A ) were treated with auxin, cultured at 37°C or 42°C for 90 min, and subjected to pull-down assays using M2 Flag beads. Co-immunoprecipitated proteins were analyzed via western blotting to detect the indicated proteins. ( C ) Predicted structural models of Claspin CKBD-AP polypeptide (aa 897–1011) in a complex with the Claspin C-terminal polypeptide (aa 1320–1339; WT, ST/A, ST/E and 5P) generated by AlphaFold3. 5P represents the peptide with 5 potential HIR/GCN2-mediated phosphorylation sites (serine and threonine) being phosphorylated. The WT and ST/A mutant could interact with the AP segment or could be accommodated into the pocket segment, whereas ST/E or the phosphorylated version of the C-terminal polypeptide is predicted not to interact with the pocket. ( D ) 293T cells were co-transfected with plasmids expressing HA-tagged Claspin C-terminal polypeptide (aa1101–1339, #28; WT, ST/A, or ST/E) and Flag-tagged Claspin AP polypeptide (aa 987–1100), as well as the plasmid expressing HRI or GCN2. Cell lysates were immunoprecipitated using anti-Flag (M2) beads, and the presence of the C-terminal polypeptide was examined.

Article Snippet: Purified Chk1 (02–117), HRI (05–154), and GCN2 (05–153) kinases were obtained from Carna Bioscience.

Techniques: Stable Transfection, Expressing, Cell Culture, Western Blot, Immunoprecipitation, Generated, Phospho-proteomics, Mutagenesis, Transfection, Plasmid Preparation

Journal: International journal of cancer

Article Title: Combined inhibition of RAD51 and CHK1 causes synergistic toxicity in cisplatin resistant cancer cells by triggering replication fork collapse.

doi: 10.1002/ijc.35164

Figure Lengend Snippet: FIGURE 2 Synergistic toxicity evoked by combined treatment with the CHK1 inhibitor PF477736 (PF) and the RAD51 inhibitor B02 in J82CisPt. (A) The viability of J82CisPt cells was analyzed after treatment with differently combined low and moderate toxic doses of CHK1i PF477736 and RAD51i B02 as indicated. Cell viability was measured after a 72 h treatment period using the AlamarBlue Assay. Based on the data obtained from three independent experiments each performed in quadruplicate, the combination indices (CIs) were calculated using CompuSyn software (CI < 0.9 indicating synergistic effects, CI ≈1 additive effects and CI > 1.2 antagonistic effects). (B) Protein expression and activation of different apoptosis-related factors were examined via Western Blot analyses using protein extracts of J82CisPt

Article Snippet: Cytostatics, DDR modifiers and other compounds were obtained from the following suppliers: Doxorubicin (Cellpharm [Bad Vilbel, Germany]), OH-Urea (Sigma [Steinheim, Germany]), 5-Fluorouracil (Medac [Wedel, Germany]), Carboplatin (TEVA [Ulm, Germany]), Oxaliplatin (Accord Healthcare [Munich, Germany]), hydrogen peroxide (H2O2) (Carl Roth GmbH [Karlsruhe, Germany]), checkpoint kinase 1 (CHK1) inhibitor PF477736 and LY2603618 (Sigma [Steinheim, Germany]), RAD51 inhibitor B02 and RI(dl)2 (Tocris Bioscience [Bristol, UK]), MRE11 inhibitor Mirin (Abcam [Cambridge, UK]), PARP inhibitor Niraparib (MedChemExpress [Monmouth Junction, NJ, USA]), HDAC inhibitors Entinostat (Selleckchem [Munich, Germany]) and Vorinostat (Sigma [Steinheim, Germany]).

Techniques: Alamar Blue Assay, Software, Expressing, Activation Assay, Western Blot

Journal: International journal of cancer

Article Title: Combined inhibition of RAD51 and CHK1 causes synergistic toxicity in cisplatin resistant cancer cells by triggering replication fork collapse.

doi: 10.1002/ijc.35164

Figure Lengend Snippet: FIGURE 3 S-phase arrest in J82CisPt cells following treatment with B02 and PF477736. (A, B) Inhibitors (10 μM B02 ± 1 μM PF477736) were added 24 h after seeding and cell cycle distribution was analyzed after a treatment period of 24 h (A) and 72 h (B) employing propidium iodide staining and flow cytometric analysis. Data are presented as mean + SD from n = 3 independent experiments. (C) The EdU incorporation of J82CisPt cells was analyzed after treatment with 10 μM B02 or/and 1 μM PF477736. EdU incorporation was analyzed after 24 h treatment period with an EdU pulse of 2 h. The graph shows the mean + SD of n = 3 independent experiments (1000–2000 nuclei analyzed per sample). The scale bars in the representative pictures correspond to 50 μm. ***p ≤.001; **p ≤.01; *p ≤.05; significant compared to control (*), to B02 mono- treatment (#) and to PF477736 mono-treatment (+). [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Cytostatics, DDR modifiers and other compounds were obtained from the following suppliers: Doxorubicin (Cellpharm [Bad Vilbel, Germany]), OH-Urea (Sigma [Steinheim, Germany]), 5-Fluorouracil (Medac [Wedel, Germany]), Carboplatin (TEVA [Ulm, Germany]), Oxaliplatin (Accord Healthcare [Munich, Germany]), hydrogen peroxide (H2O2) (Carl Roth GmbH [Karlsruhe, Germany]), checkpoint kinase 1 (CHK1) inhibitor PF477736 and LY2603618 (Sigma [Steinheim, Germany]), RAD51 inhibitor B02 and RI(dl)2 (Tocris Bioscience [Bristol, UK]), MRE11 inhibitor Mirin (Abcam [Cambridge, UK]), PARP inhibitor Niraparib (MedChemExpress [Monmouth Junction, NJ, USA]), HDAC inhibitors Entinostat (Selleckchem [Munich, Germany]) and Vorinostat (Sigma [Steinheim, Germany]).

Techniques: Staining, Control

Journal: International journal of cancer

Article Title: Combined inhibition of RAD51 and CHK1 causes synergistic toxicity in cisplatin resistant cancer cells by triggering replication fork collapse.

doi: 10.1002/ijc.35164

Figure Lengend Snippet: FIGURE 4 Hampered replication occurring after combined treatment of J82CisPt with PF477736 and B02. (A) 24 h after seeding, J82CisPt cells were treated with either 10 μM B02, 1 μM PF477736 or both substances for 6 h. After the treatment, cells were incubated for 20 min with CldU, followed by 20 min incubation with IdU. The BrdU analogs were labeled by immunofluorescence, staining was analyzed microscopically and fiber lengths were measured using ImageJ. Data presented are from two independent experiments, whereby 200 fibers were measured for each sample. Each dot represents one analyzed fiber and the black lines show the mean ± SEM. The mean value is also given above the graphs. Upper panel, nascent DNA elongation, graphically displayed as IdU track lengths of bi-colored DNA fibers. Middle panel, table summarizing the evaluation of proportions of origins and terminations in the total fiber population (ns, not significant). Lower panel, as measure of DNA replication fork stalling fork asymmetry was determined from three-colored replication origins as the ratio of the longer red IdU fiber track length versus the shorter red IdU fiber track length departing from the same green CldU track. (B) Formation of RPA foci in the nuclei of J82CisPt cells was analyzed after 6 and 24 h treatment with 10 μM B02 or/and 1 μM PF477736 via immunocytochemical staining. Data are shown with each dot representing one analyzed nucleus and the black lines showing the mean ± SEM from three independent experiments, where in each case 50 nuclei were counted. The scale bars in the representative pictures correspond to 10 μm. ***p ≤.001; **p ≤.01; *p ≤.05; significant compared to control (*), to B02 mono-treatment (#) and to PF477736 mono-treatment (+). [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Cytostatics, DDR modifiers and other compounds were obtained from the following suppliers: Doxorubicin (Cellpharm [Bad Vilbel, Germany]), OH-Urea (Sigma [Steinheim, Germany]), 5-Fluorouracil (Medac [Wedel, Germany]), Carboplatin (TEVA [Ulm, Germany]), Oxaliplatin (Accord Healthcare [Munich, Germany]), hydrogen peroxide (H2O2) (Carl Roth GmbH [Karlsruhe, Germany]), checkpoint kinase 1 (CHK1) inhibitor PF477736 and LY2603618 (Sigma [Steinheim, Germany]), RAD51 inhibitor B02 and RI(dl)2 (Tocris Bioscience [Bristol, UK]), MRE11 inhibitor Mirin (Abcam [Cambridge, UK]), PARP inhibitor Niraparib (MedChemExpress [Monmouth Junction, NJ, USA]), HDAC inhibitors Entinostat (Selleckchem [Munich, Germany]) and Vorinostat (Sigma [Steinheim, Germany]).

Techniques: Incubation, Labeling, Immunofluorescence, Staining, Control

Journal: International journal of cancer

Article Title: Combined inhibition of RAD51 and CHK1 causes synergistic toxicity in cisplatin resistant cancer cells by triggering replication fork collapse.

doi: 10.1002/ijc.35164

Figure Lengend Snippet: FIGURE 5 S-phase-dependent formation of DNA damage and activation of DDR- and DNA repair-related mechanisms following treatment of J82CisPt with PF477736 and B02. (A) Protein expression and activation of different replication stress- and DDR-related factors was examined via Western Blot analyses with protein extracts of J82CisPt cells treated for 6 h or 24 h with 10 μM B02, 1 μM PF477736 or both substances. (B) Formation of DNA strand breaks was analyzed via alkaline Comet Assay after 24 h mono- and combination-treatment with 10 μM B02 and 1 μM PF477736. Tail intensity (% DNA in tail) is displayed as dot for every analyzed cell and the mean ± SEM calculated from n = 3; N = 50. ***p ≤.001; **p ≤.01; *p ≤.05; significant compared to control (*), to B02 mono-treatment (#) and to PF477736 mono-treatment (+). (C) To analyze in which cell cycle phase the damage predominantly occurs, a double staining with γH2AX antibody and propidium iodide was applied and examined by flow cytometry after a treatment period of 6 and 24 h in J82CisPt. Displayed representative images of the flow cytometrical analyses were generated using FlowJo software. (D) J82CisPt cells were co-treated with 10 μM B02 + 1 μM PF477736 for 24 h, afterwards immunocytochemical co-staining of γH2AX and RPA was performed to analyze the correlation of both markers. For γH2AX, the mean fluorescence intensity of the nuclei was measured and the number of RPA foci per nucleus were counted. Data are shown with each dot representing one analyzed nucleus and the black lines showing the mean ± SEM from two independent experiments, where in each case 50 nuclei were measured. The scale bar in the representative picture corresponds to 20 μm. ***p ≤.001; significant compared to nuclei with <10 RPA foci. [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Cytostatics, DDR modifiers and other compounds were obtained from the following suppliers: Doxorubicin (Cellpharm [Bad Vilbel, Germany]), OH-Urea (Sigma [Steinheim, Germany]), 5-Fluorouracil (Medac [Wedel, Germany]), Carboplatin (TEVA [Ulm, Germany]), Oxaliplatin (Accord Healthcare [Munich, Germany]), hydrogen peroxide (H2O2) (Carl Roth GmbH [Karlsruhe, Germany]), checkpoint kinase 1 (CHK1) inhibitor PF477736 and LY2603618 (Sigma [Steinheim, Germany]), RAD51 inhibitor B02 and RI(dl)2 (Tocris Bioscience [Bristol, UK]), MRE11 inhibitor Mirin (Abcam [Cambridge, UK]), PARP inhibitor Niraparib (MedChemExpress [Monmouth Junction, NJ, USA]), HDAC inhibitors Entinostat (Selleckchem [Munich, Germany]) and Vorinostat (Sigma [Steinheim, Germany]).

Techniques: Activation Assay, Expressing, Western Blot, Alkaline Single Cell Gel Electrophoresis, Control, Double Staining, Flow Cytometry, Generated, Software, Staining, Fluorescence

Journal: International journal of cancer

Article Title: Combined inhibition of RAD51 and CHK1 causes synergistic toxicity in cisplatin resistant cancer cells by triggering replication fork collapse.

doi: 10.1002/ijc.35164

Figure Lengend Snippet: FIGURE 6 Combining B02 or PF477736 with other CHK1- or RAD51-inhibitors, respectively, likewise induces S-phase arrest, replication stress, and DNA damage in J82CisPt. J82CisPt cells were co-treated with 10 μM B02 + 1 μM LY2603618 (LY) (A) or 1 μM PF477736 (PF) + 30 μM RI(dl)2 (RI2) (B). Following 24 h treatment, propidium iodide-based cell cycle analysis was performed by flow cytometry with emphasis on the proportion of cells in S-phase. A total of 10,000 counts were measured for quantification. Induction of γH2AX and pRPA32 (S4, S8) was examined via Western Blot analyses with protein extracts of J82CisPt cells treated for 6 or 24 h with the corresponding combination or mono- treatments. [Color figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Cytostatics, DDR modifiers and other compounds were obtained from the following suppliers: Doxorubicin (Cellpharm [Bad Vilbel, Germany]), OH-Urea (Sigma [Steinheim, Germany]), 5-Fluorouracil (Medac [Wedel, Germany]), Carboplatin (TEVA [Ulm, Germany]), Oxaliplatin (Accord Healthcare [Munich, Germany]), hydrogen peroxide (H2O2) (Carl Roth GmbH [Karlsruhe, Germany]), checkpoint kinase 1 (CHK1) inhibitor PF477736 and LY2603618 (Sigma [Steinheim, Germany]), RAD51 inhibitor B02 and RI(dl)2 (Tocris Bioscience [Bristol, UK]), MRE11 inhibitor Mirin (Abcam [Cambridge, UK]), PARP inhibitor Niraparib (MedChemExpress [Monmouth Junction, NJ, USA]), HDAC inhibitors Entinostat (Selleckchem [Munich, Germany]) and Vorinostat (Sigma [Steinheim, Germany]).

Techniques: Cell Cycle Assay, Flow Cytometry, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Sponge-derived alkaloid AP-7 as a sensitizer to cisplatin in the treatment of multidrug-resistant NSCLC via Chk1-dependent mechanisms

doi: 10.3389/fphar.2024.1423684

Figure Lengend Snippet: DNA damage response in multidrug-resistant NSCLC cells following treatment with chemotherapy and EGFR-TKIs. Western blot analysis was performed to detect the expression of phosphorylated histone γ-H2AX, a DNA damage regulatory protein, and activated Chk1 in (A) A549/DDP cells, (B and C) PC9-GR cells, and (D) PC9-Nanog + cells after treatment with chemotherapy drugs and EGFR-TKIs. The changes in γ-H2AX levels in PC9-GR cells (E and F) and A549/DDP cells (G and H) were assessed at different time points (6, 12, 48, and 72 h) following chemotherapy treatment. (I and J) The reduction in DNA damage after 72 h of treatment with DDP or GEM compared to 12 h of treatment was also evaluated in PC9-Nanog + cells by Western blot analysis. * p < 0.05, ** p < 0.01, *** p < 0.001, compared to the control group.

Article Snippet: Firstly, enzyme, substrate, ATP and inhibitors AP-7 or the Chk1 inhibitor AZD7762 were diluted in Kinase Buffer, respectively using the CHK1 Kinase Assay Kit (Cat. V1941, Promega Corporation) according to the manufacturer’s protocol.

Techniques: Western Blot, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Sponge-derived alkaloid AP-7 as a sensitizer to cisplatin in the treatment of multidrug-resistant NSCLC via Chk1-dependent mechanisms

doi: 10.3389/fphar.2024.1423684

Figure Lengend Snippet: Combination treatment of AP-7 and DDP activates DNA damage-related signaling pathway in multidrug-resistant NSCLC cells. (A) Chk1 kinase activity was assessed by treating it with a Chk1 inhibitor and AP-7. (B,C) PC9-GR, (D,E) A549/DDP and (F,G) PC9-Nanog + cells were treated with AP-7 and DDP for 24 h, western blot was conducted to detect the protein levels of the Chk1/CDK1 pathway. *** p < 0.001, compared to the control group. * p < 0.05, ** p < 0.01, compared with DDP+AP-7 group.

Article Snippet: Firstly, enzyme, substrate, ATP and inhibitors AP-7 or the Chk1 inhibitor AZD7762 were diluted in Kinase Buffer, respectively using the CHK1 Kinase Assay Kit (Cat. V1941, Promega Corporation) according to the manufacturer’s protocol.

Techniques: Activity Assay, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Sponge-derived alkaloid AP-7 as a sensitizer to cisplatin in the treatment of multidrug-resistant NSCLC via Chk1-dependent mechanisms

doi: 10.3389/fphar.2024.1423684

Figure Lengend Snippet: AP-7 combined cisplatin inhibits the growth of gefitinib-resistant tumor in vivo . (A) Nude mice were treated with intragastric administration of 50 mg/kg gefitinib and divided into PC9 group and PC9-GR group, the volume of tumors was recorded. (B) Schedule of treatment administration is shown (n = 5, black arrows indicated the time of administration). (C) Tumor volumes of different treatment groups were measured by caliper (mean ± SD). (D) Photographs of tumor from each treatment group. (E) Immunohistochemistry results of excised tumor tissues, including Ki67, TUNEL, H&E, γ-H 2 AX, and p-Chk1 staining (×20 magnification). ### p < 0.001, compared with DDP + AP-7 group. * p < 0.05, ** p < 0.01, *** p < 0.001, compared to the ctrl group.

Article Snippet: Firstly, enzyme, substrate, ATP and inhibitors AP-7 or the Chk1 inhibitor AZD7762 were diluted in Kinase Buffer, respectively using the CHK1 Kinase Assay Kit (Cat. V1941, Promega Corporation) according to the manufacturer’s protocol.

Techniques: In Vivo, Immunohistochemistry, TUNEL Assay, Staining