anti siglec9  (R&D Systems)

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: SIGLEC9 was up regulated in cervical cancer. (A) The Protein Structures of SIGLEC9 from the Alphafold pretein structure database. (B) The expression of SIGLEC9 gene in cervical cancer from TNMplot database. (C‒D) The protein expression of SIGLEC9 was obtained from the Human Protein Atlas. (E) Association of SIGLEC9 mRNA expression and different Age, TNM and pathological stages in patients with different cancers from TCGA.

Article Snippet: Goat serum (Zhongshan Jinqiao, China) was used for blocking, followed by incubation at room temperature for 60 min. After the serum was removed, anti-CD4 (1:200, Ptroteintech, China) and SIGLEC9 (1:200, Proteintech, China) antibodies were added dropwise to the paraffin sections.

Techniques: Expressing

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Correlation analysis between SIGLEC9 and immune microenvironment in TCGA and TISIDB. (A) Violin plots comparing StromalScore, ImmuneScore, ESTIMATEScore between high and low expression of SIGLEC9, respectively. (B) The proportion of immune cell types in all patients between high- and low-SIGLEC9 groups. (C) Correlation matrix of immune cell proportions. (D) Differences in immune cell infiltration between the high and the low expression of SIGLEC9, respectively. (E) Scatter plot of the relationship between high and low SIGLEC9 expression and the level of immune cell infiltration. (F) Lollipop plot of SIGLEC9 expression in relation to immune cells. (G) Spearman correlations between expression of SIGLEC9 and TILs across CESC. (H) The TISIDB database to analyzed SIGLEC9 expression in macrophages, MDSC, Act DC and Treg cells. (I) SIGLEC9 mRNA expression in different immune subtypes in CESC via TISIDB. (J) Relationship between SIGLEC9 expression and immune checkpoints. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Goat serum (Zhongshan Jinqiao, China) was used for blocking, followed by incubation at room temperature for 60 min. After the serum was removed, anti-CD4 (1:200, Ptroteintech, China) and SIGLEC9 (1:200, Proteintech, China) antibodies were added dropwise to the paraffin sections.

Techniques: Expressing

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Differential expression analysis and cell subset identification in cervical cancer using single-cell analysis. (A‒B) Stringent cell filtration criteria were applied, including the selection of cells with a gene count between 200 and 5000 (nFeature > 200 and ≤ 5000) and a limited proportion of mitochondrial genes (percent.mt < 10). (C‒F) Principal Component Analysis was performed. (G) Expression profiles of well-known markers were assessed across different cell types in the central nervous system (CC). (H‒N) Bubble plot of marker gene expression in the identification of different cell types. (O‒P) The uniform manifold approximation and projection and the t-distributed Stochastic Neighbor Embedding (t-SNE) plot demonstrating main cell types in CC. (Q‒R) Exhibition of group (nc = normal, rm = tumor). (S‒T) Exhibition of cell subsets. (U) Distribution of cells in each sample. (V) Comparison of SIGLEC9 expression between tumor tissue and adjacent normal tissue. (W) Differential expression of SIGLEC9 between tumor tissue and normal tissue. (X1) The UMAP of macrophages. (X2) Macrophages into two groups with high and low SIGLEC9 expression. (X3) DEGs between high- and low-SIGLEC9 groups in macrophages. (Y1‒2) Gene Ontology analysis between high- and low-SIGLEC9 groups in macrophages. (Y3) Kyoto Encyclopedia of Genes and Genomes analysis between high- and low-SIGLEC9 groups in macrophages.

Article Snippet: Goat serum (Zhongshan Jinqiao, China) was used for blocking, followed by incubation at room temperature for 60 min. After the serum was removed, anti-CD4 (1:200, Ptroteintech, China) and SIGLEC9 (1:200, Proteintech, China) antibodies were added dropwise to the paraffin sections.

Techniques: Quantitative Proteomics, Single-cell Analysis, Filtration, Selection, Expressing, Marker, Gene Expression, Comparison

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: PPI map of proteins interacting with SIGLEC9. (A‒D) The PPI network of SIGLEC9 from STRING (A), IntAct (B), BioGIRD (C), Mentha (D). (E) The expression of MUC1 gene in cervical cancer and normal cervical tissue from GEO database. (F) Expression of MUC1 gene in cervical cancer with or without lymph node metastasis from GEO database. (G) The expression of MUC1 gene in cervical cancer and normal cervical tissue from GEPIA database (* p < 0.05).

Article Snippet: Goat serum (Zhongshan Jinqiao, China) was used for blocking, followed by incubation at room temperature for 60 min. After the serum was removed, anti-CD4 (1:200, Ptroteintech, China) and SIGLEC9 (1:200, Proteintech, China) antibodies were added dropwise to the paraffin sections.

Techniques: Expressing

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Expression level and clinical correlation in cervical cancer. (A) The expression of SIGLEC9 in cervical cancer patients ( n = 58) and normal control ( n = 30) with immunohistochemical staining. (B) The expression of SIGLEC9 in cervical cancer patients with western blotting ( n = 8). (C) The cervical cancer patients with a high SIGLEC9 expression ( n = 29) had a shorter survival probability than those patients with a low SIGLEC9 expression ( n = 29) ( p = 0.0057). * p < 0.05, ** p < 0.01.

Article Snippet: Goat serum (Zhongshan Jinqiao, China) was used for blocking, followed by incubation at room temperature for 60 min. After the serum was removed, anti-CD4 (1:200, Ptroteintech, China) and SIGLEC9 (1:200, Proteintech, China) antibodies were added dropwise to the paraffin sections.

Techniques: Expressing, Control, Immunohistochemical staining, Staining, Western Blot

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Interaction of SIGLEC9 with CD4+ T-cell and MUC1. (A) Immunohistochemical expression of MUC1 in cervical cancer and normal cervical tissues ( n = 40). (B) MUC1 protein expression was analyzed by cell immunofluorescence ( n = 3, three independent experiments). (C) The expression of SIGLEC9 (Green) and CD4 (Red), DAPI (Blue) in normal cervical tissue and in cancer tissues with double immunofluorescence. (D) Correlation between MUC1 (red) and SIGLEC9 (green), DAPI (Blue) in normal tissues ( n = 6) and cancer tissues ( n = 6). (E‒F) Multiplexed immunofluorescence for SIGLEC9 (Red), CD4 (Yellow), and MUC1 (Green), and DAPI (Blue) in cancer tissues and normal tissues. (G‒I) The flow cytometry expression of SIGLEC9+ CD4+ T/CD8+ T-cells and SIGLEC9+M1/M2 in cervical cancer patients ( n = 40) and normal control ( n = 20). ** p < 0.01, *** p < 0.001, and **** p < 0.0001. (J) The cervical cancer patients with a high SIGLEC9+ TAM cell infiltration ( n = 20) had a shorter survival probability than those patients with a low SIGLEC9+ TAM cell infiltration ( n = 20; p = 0.0049).

Article Snippet: Goat serum (Zhongshan Jinqiao, China) was used for blocking, followed by incubation at room temperature for 60 min. After the serum was removed, anti-CD4 (1:200, Ptroteintech, China) and SIGLEC9 (1:200, Proteintech, China) antibodies were added dropwise to the paraffin sections.

Techniques: Immunohistochemical staining, Expressing, Immunofluorescence, Flow Cytometry, Control

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: SIGLEC9 was up regulated in cervical cancer. (A) The Protein Structures of SIGLEC9 from the Alphafold pretein structure database. (B) The expression of SIGLEC9 gene in cervical cancer from TNMplot database. (C‒D) The protein expression of SIGLEC9 was obtained from the Human Protein Atlas. (E) Association of SIGLEC9 mRNA expression and different Age, TNM and pathological stages in patients with different cancers from TCGA.

Article Snippet: The protein samples were then heated at 100 °C for 10 min. SDS-PAGE electrophoresis was performed at 10 % concentration for 2 h, followed by membrane transfer onto PVDF membranes (Millipore, Germany) for 1 h. The membranes were incubated with 5 % skim milk for 1 h. Subsequently, membranes were incubated overnight at 4 °C with primary antibodies: rabbit anti-human SIGLEC9 (1:1000, Proteintech, Chinav) and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Correlation analysis between SIGLEC9 and immune microenvironment in TCGA and TISIDB. (A) Violin plots comparing StromalScore, ImmuneScore, ESTIMATEScore between high and low expression of SIGLEC9, respectively. (B) The proportion of immune cell types in all patients between high- and low-SIGLEC9 groups. (C) Correlation matrix of immune cell proportions. (D) Differences in immune cell infiltration between the high and the low expression of SIGLEC9, respectively. (E) Scatter plot of the relationship between high and low SIGLEC9 expression and the level of immune cell infiltration. (F) Lollipop plot of SIGLEC9 expression in relation to immune cells. (G) Spearman correlations between expression of SIGLEC9 and TILs across CESC. (H) The TISIDB database to analyzed SIGLEC9 expression in macrophages, MDSC, Act DC and Treg cells. (I) SIGLEC9 mRNA expression in different immune subtypes in CESC via TISIDB. (J) Relationship between SIGLEC9 expression and immune checkpoints. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: The protein samples were then heated at 100 °C for 10 min. SDS-PAGE electrophoresis was performed at 10 % concentration for 2 h, followed by membrane transfer onto PVDF membranes (Millipore, Germany) for 1 h. The membranes were incubated with 5 % skim milk for 1 h. Subsequently, membranes were incubated overnight at 4 °C with primary antibodies: rabbit anti-human SIGLEC9 (1:1000, Proteintech, Chinav) and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Differential expression analysis and cell subset identification in cervical cancer using single-cell analysis. (A‒B) Stringent cell filtration criteria were applied, including the selection of cells with a gene count between 200 and 5000 (nFeature > 200 and ≤ 5000) and a limited proportion of mitochondrial genes (percent.mt < 10). (C‒F) Principal Component Analysis was performed. (G) Expression profiles of well-known markers were assessed across different cell types in the central nervous system (CC). (H‒N) Bubble plot of marker gene expression in the identification of different cell types. (O‒P) The uniform manifold approximation and projection and the t-distributed Stochastic Neighbor Embedding (t-SNE) plot demonstrating main cell types in CC. (Q‒R) Exhibition of group (nc = normal, rm = tumor). (S‒T) Exhibition of cell subsets. (U) Distribution of cells in each sample. (V) Comparison of SIGLEC9 expression between tumor tissue and adjacent normal tissue. (W) Differential expression of SIGLEC9 between tumor tissue and normal tissue. (X1) The UMAP of macrophages. (X2) Macrophages into two groups with high and low SIGLEC9 expression. (X3) DEGs between high- and low-SIGLEC9 groups in macrophages. (Y1‒2) Gene Ontology analysis between high- and low-SIGLEC9 groups in macrophages. (Y3) Kyoto Encyclopedia of Genes and Genomes analysis between high- and low-SIGLEC9 groups in macrophages.

Article Snippet: The protein samples were then heated at 100 °C for 10 min. SDS-PAGE electrophoresis was performed at 10 % concentration for 2 h, followed by membrane transfer onto PVDF membranes (Millipore, Germany) for 1 h. The membranes were incubated with 5 % skim milk for 1 h. Subsequently, membranes were incubated overnight at 4 °C with primary antibodies: rabbit anti-human SIGLEC9 (1:1000, Proteintech, Chinav) and GAPDH (1:3000, Proteintech, China).

Techniques: Quantitative Proteomics, Single-cell Analysis, Filtration, Selection, Expressing, Marker, Gene Expression, Comparison

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: PPI map of proteins interacting with SIGLEC9. (A‒D) The PPI network of SIGLEC9 from STRING (A), IntAct (B), BioGIRD (C), Mentha (D). (E) The expression of MUC1 gene in cervical cancer and normal cervical tissue from GEO database. (F) Expression of MUC1 gene in cervical cancer with or without lymph node metastasis from GEO database. (G) The expression of MUC1 gene in cervical cancer and normal cervical tissue from GEPIA database (* p < 0.05).

Article Snippet: The protein samples were then heated at 100 °C for 10 min. SDS-PAGE electrophoresis was performed at 10 % concentration for 2 h, followed by membrane transfer onto PVDF membranes (Millipore, Germany) for 1 h. The membranes were incubated with 5 % skim milk for 1 h. Subsequently, membranes were incubated overnight at 4 °C with primary antibodies: rabbit anti-human SIGLEC9 (1:1000, Proteintech, Chinav) and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Expression level and clinical correlation in cervical cancer. (A) The expression of SIGLEC9 in cervical cancer patients ( n = 58) and normal control ( n = 30) with immunohistochemical staining. (B) The expression of SIGLEC9 in cervical cancer patients with western blotting ( n = 8). (C) The cervical cancer patients with a high SIGLEC9 expression ( n = 29) had a shorter survival probability than those patients with a low SIGLEC9 expression ( n = 29) ( p = 0.0057). * p < 0.05, ** p < 0.01.

Article Snippet: The protein samples were then heated at 100 °C for 10 min. SDS-PAGE electrophoresis was performed at 10 % concentration for 2 h, followed by membrane transfer onto PVDF membranes (Millipore, Germany) for 1 h. The membranes were incubated with 5 % skim milk for 1 h. Subsequently, membranes were incubated overnight at 4 °C with primary antibodies: rabbit anti-human SIGLEC9 (1:1000, Proteintech, Chinav) and GAPDH (1:3000, Proteintech, China).

Techniques: Expressing, Control, Immunohistochemical staining, Staining, Western Blot

Journal: Clinics

Article Title: The role of SIGLEC9 in immunosuppression and prognosis in cervical cancer

doi: 10.1016/j.clinsp.2025.100849

Figure Lengend Snippet: Interaction of SIGLEC9 with CD4+ T-cell and MUC1. (A) Immunohistochemical expression of MUC1 in cervical cancer and normal cervical tissues ( n = 40). (B) MUC1 protein expression was analyzed by cell immunofluorescence ( n = 3, three independent experiments). (C) The expression of SIGLEC9 (Green) and CD4 (Red), DAPI (Blue) in normal cervical tissue and in cancer tissues with double immunofluorescence. (D) Correlation between MUC1 (red) and SIGLEC9 (green), DAPI (Blue) in normal tissues ( n = 6) and cancer tissues ( n = 6). (E‒F) Multiplexed immunofluorescence for SIGLEC9 (Red), CD4 (Yellow), and MUC1 (Green), and DAPI (Blue) in cancer tissues and normal tissues. (G‒I) The flow cytometry expression of SIGLEC9+ CD4+ T/CD8+ T-cells and SIGLEC9+M1/M2 in cervical cancer patients ( n = 40) and normal control ( n = 20). ** p < 0.01, *** p < 0.001, and **** p < 0.0001. (J) The cervical cancer patients with a high SIGLEC9+ TAM cell infiltration ( n = 20) had a shorter survival probability than those patients with a low SIGLEC9+ TAM cell infiltration ( n = 20; p = 0.0049).

Article Snippet: The protein samples were then heated at 100 °C for 10 min. SDS-PAGE electrophoresis was performed at 10 % concentration for 2 h, followed by membrane transfer onto PVDF membranes (Millipore, Germany) for 1 h. The membranes were incubated with 5 % skim milk for 1 h. Subsequently, membranes were incubated overnight at 4 °C with primary antibodies: rabbit anti-human SIGLEC9 (1:1000, Proteintech, Chinav) and GAPDH (1:3000, Proteintech, China).

Techniques: Immunohistochemical staining, Expressing, Immunofluorescence, Flow Cytometry, Control