human gal9 protein  (R&D Systems)

Journal: Molecular Cancer

Article Title: Targeting the CD47-HCK-LGALS9 axis disrupts proliferation-immunosuppression coupling in early-stage endometrial cancer

doi: 10.1186/s12943-025-02534-0

Figure Lengend Snippet: Cell-cell communication analysis and spatial localization of the oncogenic subpopulation. A Chord diagram showing the number of interactions between epithelial cells and myeloid-derived cells, where line thickness indicates the interaction count. B Heatmap depicting the interaction strength between epithelial cells and myeloid-derived cells. C Dotplot of receptor-ligand pairs in intercellular communication between epithelial cells and myeloid-derived cells, with red circles representing P < 0.05. D The expression of LGALS9 and SIRPA in macrophages in tumor and normal samples. E Distribution of CD47 expression levels across all cells along the pseudotime axis, with the color gradient (blue to red) representing low to high expression. F , G Scatter plot showing the correlation between pseudotime progression and CD47 ( F ) and CDK1 ( G ) expression levels. H Spatial localization of CD47 and CDK1. I Spatial localization of cell subpopulations. Significance levels: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HEC-1 A and KLE cells were treated with the following agents: RRX-001 (#HY-16438, MCE, USA) at 5, 6.25, and 7.5 μM; recombinant LGALS9 (#11147-H01H, Sino Biological, China) at 50 ng/mL; GSK5182 (#SML3150, Germany) at 10, 20, and 30 μM; progesterone (#HY-N0437, MCE, USA); and estradiol (#HY-B0141, MCE, USA).

Techniques: Derivative Assay, Expressing

Journal: Molecular Cancer

Article Title: Targeting the CD47-HCK-LGALS9 axis disrupts proliferation-immunosuppression coupling in early-stage endometrial cancer

doi: 10.1186/s12943-025-02534-0

Figure Lengend Snippet: The interaction of the LGALS9-CD47 axis and the formation of an immunosuppressive microenvironment in the co-culture model of EC cells and macrophages. A Molecular docking predicting the interaction between LGALS9 and CD47. B The co-culture model of EC cells and macrophages. C ELISA analysis of LGALS9 secretion levels in the supernatant of macrophages under different conditions: EC cells, macrophages (M), EC cells + M, and EC cells + M + RRX-001. D , E ELISA analysis of IL-10 and TGF-β1 secretion levels in the supernatant of macrophages under different conditions: M, CD47 + CDK1 + EC cells + M, and CD47 − CDK1 − EC cells + M. F Represents the phagocytic images of macrophages incubated with GFP-labeled EC cells, with a scale bar of 50 μm; G Depicts the FACS-based phagocytosis diagram, where Q1 represents unphagocytosed GFP + EC cells, Q2 represents phagocytosed GFP + CD45 + macrophages, and Q3 represents unphagocytosed CD45 + macrophages. H WB results of GST-pull down MS. Input represents the whole cell lysate before the experiment, used to confirm the expression of the bait protein GST-CD47 and target proteins. Lanes: 1, GST; 2, GST-CD47; 3, GST + macrophage sample; 4, GST-CD47 + macrophage sample. I Silver staining results of GST-pull down MS. Control1 represents GST + sample, Control2 represents GST-CD47 + lysate, and Sample represents GST-CD47 + macrophage sample. J Changes in membrane protein expression levels in the co-culture model. K Molecular docking predicting the interaction between HCK and CD47. Significance levels: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HEC-1 A and KLE cells were treated with the following agents: RRX-001 (#HY-16438, MCE, USA) at 5, 6.25, and 7.5 μM; recombinant LGALS9 (#11147-H01H, Sino Biological, China) at 50 ng/mL; GSK5182 (#SML3150, Germany) at 10, 20, and 30 μM; progesterone (#HY-N0437, MCE, USA); and estradiol (#HY-B0141, MCE, USA).

Techniques: Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Incubation, Labeling, Expressing, Silver Staining, Membrane

Journal: Molecular Cancer

Article Title: Targeting the CD47-HCK-LGALS9 axis disrupts proliferation-immunosuppression coupling in early-stage endometrial cancer

doi: 10.1186/s12943-025-02534-0

Figure Lengend Snippet: The LGALS9-CD47 axis promotes EC cell proliferation. A , B . Changes in cell viability after 24 h and 5 days of treatment with rLGALS9 and RRX-001, detected by the CCK-8 assay. C - F . Effects of rLGALS9 and CD47 inhibitor treatment for 24 h on cell apoptosis and cell cycle, analyzed by flow cytometry. G Expression levels of CD47, cyclin D1 and PCNA at the mRNA levels in EC cells treated with rLGALS9 and RRX-001 for 24 h. H , I After treating with rLGALS9 and RRX-001 for 24 h in EC cells, the expression of CD47, cyclin D1, PCNA, Bcl-2 and Bax was measured by WB, with GAPDH used as a loading control. Significance levels: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HEC-1 A and KLE cells were treated with the following agents: RRX-001 (#HY-16438, MCE, USA) at 5, 6.25, and 7.5 μM; recombinant LGALS9 (#11147-H01H, Sino Biological, China) at 50 ng/mL; GSK5182 (#SML3150, Germany) at 10, 20, and 30 μM; progesterone (#HY-N0437, MCE, USA); and estradiol (#HY-B0141, MCE, USA).

Techniques: CCK-8 Assay, Flow Cytometry, Expressing, Control

Journal: Molecular Cancer

Article Title: Targeting the CD47-HCK-LGALS9 axis disrupts proliferation-immunosuppression coupling in early-stage endometrial cancer

doi: 10.1186/s12943-025-02534-0

Figure Lengend Snippet: Correlation between CD47 and EC in clinical samples. A , B . Immunofluorescence was performed to identify CD47 + CDK1 + positive cells in normal and EC epithelium (3 ROIs per group). Scale bar = 100 μm ( A ). Bar chart of the proportion of CD47 + CDK1 + cells in tumor epithelium and adjacent normal epithelial ROI ( B ). C - E . Representative images of the IHC of CD47 and LGALS9 in cancer tissues and normal tissues and a statistical plot of CD47 and LGALS9 expression in different groups. F Brief flow chart of proteomics. G . Expression of CD47 in 24 tumor samples and adjacent normal endometrial tissues. H . The expression of CD47 in 15 cases of favorable prognosis and 9 cases of poor prognosis EC tissues. I . Prediction of survival changes in EC patients with different expression levels of CD47 using proteomics data. EC patients were stratified into low-risk and high-risk groups based on the median expression value. Survival curves were compared using the Log-rank test and visualized with the Kaplan-Meier method. Significance levels: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HEC-1 A and KLE cells were treated with the following agents: RRX-001 (#HY-16438, MCE, USA) at 5, 6.25, and 7.5 μM; recombinant LGALS9 (#11147-H01H, Sino Biological, China) at 50 ng/mL; GSK5182 (#SML3150, Germany) at 10, 20, and 30 μM; progesterone (#HY-N0437, MCE, USA); and estradiol (#HY-B0141, MCE, USA).

Techniques: Immunofluorescence, Expressing

Journal: Molecular Cancer

Article Title: Targeting the CD47-HCK-LGALS9 axis disrupts proliferation-immunosuppression coupling in early-stage endometrial cancer

doi: 10.1186/s12943-025-02534-0

Figure Lengend Snippet: The role of the LGALS9-CD47 axis in early-stage EC was confirmed in PDOs. A Flow cytometric analysis of CD47 expression in PDOs transfected with ov-CD47 virus ( n = 3). B Bar graph showing the expression levels of CD47 in PDOs transfected with control and ov-CD47 virus. C The morphological changes in PDOs at 0 h, 72 h, and 120 h were evaluated using light microscopy after transfection with control and ov-CD47 virus ( n = 3). D The cell viability treated with different viruses was evaluated by CCL 3D with three replications. E , F Phagocytosis of indicated PDOs was represented by the percentage of FITC + CD14 + cells in total CD14 + cells ( n = 3). G - I . The levels of LGALS9, IL-10 and TGF-β1 secreted by PBMC-derived macrophages (M) were detected by ELISA. J The morphological changes in PDOs at 0 h, 24 h, and 72 h were evaluated using light microscopy with or without rLGALS9 treatment ( n = 3). K . The cell viability treated with different treatments was evaluated by CCL 3D with three replications. Significance levels: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: HEC-1 A and KLE cells were treated with the following agents: RRX-001 (#HY-16438, MCE, USA) at 5, 6.25, and 7.5 μM; recombinant LGALS9 (#11147-H01H, Sino Biological, China) at 50 ng/mL; GSK5182 (#SML3150, Germany) at 10, 20, and 30 μM; progesterone (#HY-N0437, MCE, USA); and estradiol (#HY-B0141, MCE, USA).

Techniques: Expressing, Transfection, Virus, Control, Light Microscopy, Derivative Assay, Enzyme-linked Immunosorbent Assay