anti checkpoint kinase 1  (MedChemExpress)

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: Molecular Medicine

Article Title: Activation of the MEK1-CHK2 axis in macrophages by Staphylococcus aureus promotes mitophagy, resulting in a reduction in bactericidal efficacy

doi: 10.1186/s10020-025-01274-7

Figure Lengend Snippet: MEK1-ERK1/2 activation in macrophages enhances mitophagy by inhibiting CHEK2 expression to reduce mtROS levels. A – D Representative images ( A ) and quantification ( B – D ) of CHEK2 levels in BMDMs infected with S. aureus (MOI = 10) for 12 h and treated with or without GDC-0973 or LY3214996. Statistical results of CHEK2 fluorescence intensity ( B ), mitochondrial localization of CHEK2 ( C ), and nuclear localization of CHEK2 ( D ). Scale bars, 10 μm. E and F Representative images ( E ) and quantification ( F ) of mtROS levels in BMDMs infected with S. aureus for 12 h and treated with GDC-0973 or LY3214996 and BML-277. Scale bars, 5 μm. G – I Representative images ( G ) and quantification ( H and I ) of LysoTracker/MitoTracker staining in BMDMs infected with S. aureus (MOI = 10) for 12 h and treated with GDC-0973 or LY3214996 and BML-277. Scale bars, 10 μm. n = 5 samples/group, *** p < 0.001

Article Snippet: Following PBS washes, the cells were incubated in blocking buffer at room temperature in the dark for 1 h and then incubated with a rabbit anti-checkpoint kinase 2 (CHEK2) primary antibody (#13954-1-AP, Proteintech, USA) for 2 h at room temperature in the dark.

Techniques: Activation Assay, Expressing, Infection, Fluorescence, Staining

Journal: Molecular Medicine

Article Title: Activation of the MEK1-CHK2 axis in macrophages by Staphylococcus aureus promotes mitophagy, resulting in a reduction in bactericidal efficacy

doi: 10.1186/s10020-025-01274-7

Figure Lengend Snippet: Mitophagy in macrophages regulated by the MEK1-ERK1/2-CHEK2 signaling pathway is crucial for the progression of S. aureus -induced osteomyelitis. A and B Representative images ( A ) and quantitative ( B ) analysis of H&E staining of S. aureus -infected femurs from mice treated with or without GDC-0973 or LY3214996 and BML-277 compared with those treated with the vehicle. Scale bars, 100 μm. C and D Representative images ( C ) and quantitative ( D ) analysis of the bacterial load in the S. aureus -infected femurs of mice treated with or without GDC-0973 or LY3214996 and BML-277 compared with those treated with the vehicle. E and F Representative images ( E ) and quantification ( F ) of the flow cytometry results for the proportions of LysoTracker + Mitophagy+/F4/80 + CD11b + cells in the S. aureus -infected femurs of mice treated with or without GDC-0973 or LY3214996 and BML-277 compared with those treated with the vehicle. n = 5 mice/group, *** p < 0.001

Article Snippet: Following PBS washes, the cells were incubated in blocking buffer at room temperature in the dark for 1 h and then incubated with a rabbit anti-checkpoint kinase 2 (CHEK2) primary antibody (#13954-1-AP, Proteintech, USA) for 2 h at room temperature in the dark.

Techniques: Staining, Infection, Flow Cytometry