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

Journal: The Journal of Clinical Investigation

Article Title: Organ-specific features of human kidney lymphatics are disrupted in chronic transplant rejection

doi: 10.1172/JCI168962

Figure Lengend Snippet: ( A ) Violin plots showing upregulation of IFN-inducible genes IFITM2 and IFITM3 in LECs from rejecting allografts. ( B ) UMAP of the scRNA-Seq data showing enrichment of IFN-γ within the T/NK cell cluster. ( C ) UMAP showing enrichment of an IFN-γ signature, including IFNGR1 , IFNGR2 , IFITM2 , and IFITM3 . ( D ) CellPhoneDB interaction map depicting predicted lymphatic-CD4 + T cell crosstalk in rejection. Inhibitory interactions (blue) include PVR and LGALS9; stimulatory interactions (red) are also shown. Node size reflects expression frequency; line intensity indicates interaction strength. Ligands of interest, PVR and LGALS9 , are highlighted. ( E ) Heatmap of immune checkpoint interactions between LECs and effector CD4 + T cells across disease states. Color indicates normalized CellPhoneDB interaction score. All scores were normalized for each ligand-receptor pair. ( F ) Immunofluorescence validation of PVR expression on PDPN + lymphatics (arrowhead) in rejecting kidneys ( n = 2); CD4 + T cell shown in contact (asterisk). Scale bar: 30 μm. ( G ) IFN-γ stimulation of cultured human LECs increases LGALS9 levels at 24 and 48 hours (qPCR; *** P = 0.0002, ** P = 0.0093, respectively) relative to HPRT. ( H ) LGALS9 protein secretion increased at 48 and 72 hours (ELISA; *** P = 0.0002, **** P < 0.0001, respectively) after IFN-γ stimulation of cultured human LECs. qPCR and ELISA experiments were repeated 3 times, and all assays were performed in duplicate, with each dot on the graph representing the mean data obtained for each repeat.

Article Snippet: Secreted LGALS9 protein in conditioned media was measured by ELISA (R&D Systems).

Techniques: Expressing, Immunofluorescence, Biomarker Discovery, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Myeloid‐Derived LGALS9 ‐ P4HB Immune Interaction Promotes Metastasis in Gastric Cancer Through Enhanced Cell Proliferation and Lipid Metabolism

doi: 10.1111/jcmm.70661

Figure Lengend Snippet: Cell–cell communication analysis reveals LGALS9–P4HB as a key interaction between myeloid and epithelial cells in metastatic sites. (A) Heatmap showing total cell interactions in liver metastases. (B) Heatmap showing total cell interactions in lymph node metastases. (C) Outgoing and incoming signalling patterns in liver metastases. (D) Outgoing and incoming signalling patterns in lymph node metastases. (E) Dot plot showing ligand–receptor pairs mediating immune cell–epithelial cell communication in liver metastases. (F) Dot plot showing ligand–receptor pairs mediating immune cell–epithelial cell communication in lymph node metastases. (G) Network diagram of cellular interactions in liver metastases. (H) Network diagram of cellular interactions in lymph node metastases. (I) Violin plots showing expression of LGALS9 and its receptors (P4HB, HAVCR2, CD44) across different cell types in liver metastases. (J) Violin plots showing expression of LGALS9 and its receptors across different cell types in lymph node metastases.

Article Snippet: Recombinant human LGALS9 protein (R&D Systems) was added to the culture medium at a concentration of 150 ng/mL.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Myeloid‐Derived LGALS9 ‐ P4HB Immune Interaction Promotes Metastasis in Gastric Cancer Through Enhanced Cell Proliferation and Lipid Metabolism

doi: 10.1111/jcmm.70661

Figure Lengend Snippet: LGALS9 activates P4HB to promote gastric cancer cell proliferation, EMT and lipid metabolism. (A) Relative expression of P4HB in different gastric cell lines measured by qRT‐PCR. (B) Expression of proliferation‐related genes (Ki‐67, PCNA, Cyclin D1) in MKN‐74 cells treated with recombinant LGALS9 or LGALS9 conditional medium, determined by qRT‐PCR. (C) Cell viability of MKN‐74 cells treated with recombinant LGALS9 over time, measured by MTT assay. (D) Cell viability of MKN‐74 cells cultured in LGALS9 conditional medium over time. (E) EdU incorporation assay showing proliferation of MKN‐74 cells treated with recombinant LGALS9. (F) EdU incorporation assay showing proliferation of MKN‐74 cells cultured in LGALS9 conditional medium. (G) Western blot analysis of EMT markers (E‐cadherin, N‐cadherin, Vimentin) in MKN‐74 cells with indicated treatments. (H) qRT‐PCR analysis of lipid metabolism–related genes in MKN‐74 cells treated with LGALS9 conditional medium (upper panel) or recombinant LGALS9 (lower panel). Statistical significance is indicated by asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001, ns: Not significant.

Article Snippet: Recombinant human LGALS9 protein (R&D Systems) was added to the culture medium at a concentration of 150 ng/mL.

Techniques: Expressing, Quantitative RT-PCR, Recombinant, MTT Assay, Cell Culture, Western Blot