mouse anti human spp1 monoclonal antibody  (Santa Cruz Biotechnology)

Journal: Experimental & Molecular Medicine

Article Title: Single-cell transcriptomics of the myeloid milieu reveals an angiogenic niche in triple-negative breast cancer

doi: 10.1038/s12276-025-01571-5

Figure Lengend Snippet: a UMAP of the monocytes and macrophages in TNBCs. The pie chart presents the proportions of each subtype. b Heatmap depicting the expression of the DEGs across the monocyte and macrophage subtypes. c Expression in all subtypes of the genes that mark each subtype. The bar plot displays the macrophage differentiation score (Mφ Diff), which was computed using AUCell on the basis of the gene set GO: 0030225. The dashed line indicates the mean AUC score of the subtypes. The monocyte subtypes had lower differentiation scores, whereas all macrophage subtypes apart from SPP1 hi Mφs and MKI67 hi Mφs had scores above the mean. d Two-dimensional visualization of the differentiation trajectories of the monocytes and monocyte-derived macrophages, as determined with a diffusion-map approach. The arrows indicate the trajectories that were calculated with slingshot. They all start from VCAN hi Monos but end with either SPP1 hi Mφs, IL1B hi Mφs or CCL3 hi Mφs. e Estimated differentiation trajectory of monocytes and monocyte-derived macrophages, as determined with scTour. These data are displayed in a vector map (top) and as pseudotime scores (bottom). The lineage that results in SPP1 hi Mφs is distinct from the classical lineage that involves the differentiation of IL1B hi Mφs into CCL3 hi Mφs. f GSEA comparing the classical lineage and SPP1 lineage. The classical lineage is marked by enhanced inflammation-related pathways while the SPP1 lineage displays increased metabolic stress-related pathway activity. g The GSVA-calculated pathway activities in the monocyte/macrophage subtypes. The color and size of each dot indicate the log 2 fold change and adjusted P value, respectively. SPP1hi Mφs are enriched in glycolysis, hypoxia and tumor vasculature pathways. h Kaplan–Meier plot depicting the OS of patients with TNBC in the TCGA cohort after their stratification according to their high (red) or low (black) expression of SPP1 hi Mφ markers. The P value was calculated using Cox regression. HR, hazard ratio; CI, confidence interval.

Article Snippet: The primary antibodies were as follows: SPP1 (HPA027541; Atlas; 1:300), ADM (TA806874; Origene; 1:2,000), VEGF (sc-7269; Santa Cruz; 1:4,000); CD68 (ab955; abcam; 1:12,000); CD66b (NB1000-77808; Novus; 1:400), Apelin (ab230536; abcam; 1:50).

Techniques: Expressing, Derivative Assay, Diffusion-based Assay, Plasmid Preparation, Activity Assay

Journal: Experimental & Molecular Medicine

Article Title: Single-cell transcriptomics of the myeloid milieu reveals an angiogenic niche in triple-negative breast cancer

doi: 10.1038/s12276-025-01571-5

Figure Lengend Snippet: a A schematic depiction of the spatial transcriptomic analysis performed in the present study. Our TNBC myeloid cell scRNA-seq atlas was combined with the publicly available Visium data on TNBC samples ( GSE210616 ) . b The estimated proportion of each macrophage subtype in the total macrophage population of each Visium sample. Abundance was calculated using cell2location. c Correlation between the macrophage subtype proportions across the Visium spots. SPP1 hi Mφs correlated poorly with the other subtypes. d , e The spatial distribution of the macrophage subtypes and epithelial cancer cells in the representative Visium slides denoted 094D ( d ) and 395D ( e ). The hypoxia-pathway activity in each Visium spot was also calculated using decoupler. SPP1 hi Mφs colocalized with epithelial cells in hypoxic regions. f Correlation between the transcriptomic signatures of various pathways from the MSigDB database and the macrophage subtype proportions across Visium spots. SPP1 hi Mφs show heightened hypoxia-pathway activity. g Cell–cell interaction from macrophage subtypes to epithelial cells, as inferred by CellChat analysis. h Cell–cell interaction from epithelial cells to macrophage subtypes, as inferred by CellChat analysis. i Spatial-expression patterns of the epithelial-ligand MIF and its CD44 receptor on SPP1 hi Mφs in 094D, a representative slide. High expression of CD44 and MIF was observed in the hypoxic niches. *P < 0.05.

Article Snippet: The primary antibodies were as follows: SPP1 (HPA027541; Atlas; 1:300), ADM (TA806874; Origene; 1:2,000), VEGF (sc-7269; Santa Cruz; 1:4,000); CD68 (ab955; abcam; 1:12,000); CD66b (NB1000-77808; Novus; 1:400), Apelin (ab230536; abcam; 1:50).

Techniques: Activity Assay, Expressing

Journal: Experimental & Molecular Medicine

Article Title: Single-cell transcriptomics of the myeloid milieu reveals an angiogenic niche in triple-negative breast cancer

doi: 10.1038/s12276-025-01571-5

Figure Lengend Snippet: a UMAP of the intratumoral EC subtypes and their APLN expression. Dot size and color of the dotplot correspond to mean expression and fraction of cells, respectively. APLN was only expressed by the CapEC 1 subset. b A matrix plot displaying the correlation between the macrophage subtypes and EC markers. SPP1hi Mφs show negative or no correlation with SEMA3G , ACKR1 and RGCC but correlate strongly with VEGFA and APLN . c A representative slide (094D) showing the expression of VEGFA and APLN and the distribution of SPP1 hi Mφs and VEGFA hi neutrophils. The SPP1 hi Mφs and VEGFA hi neutrophils colocalized in the region that bears high VEGFA and APLN expression. d A matrix plot depicting the major ligand–receptor pairs between the macrophage subtypes and the three capillary EC subtypes. Of the macrophage subtypes, SPP1 hi Mφs interact most strongly with CapEC 1 . e ADM expression in the monocytes and macrophages in our atlas. ADM expression localizes to the SPP1 hi monocytes and the SPP1 hi Mφs. f Spatial correlation between macrophage subtypes and ADM expression, as shown by a matrix plot. g mIF results showing the presence of an angiogenic niche a patient sample. The colocalization of SPP1 hi Mφs and VEGFA hi neutrophils with APLN-expressing cells can be seen. Scale bars, 200 µm (top) and 50 µm (bottom). h Hypoxia induces THP-1 cells into SPP1 hi Mφ-like cells. PMA-treated THP-1 cells were cultured in hypoxic conditions for 24 h. Expression of SPP1 and ADM was validated by RT–qPCR. The SPP1 protein level was measured by western blot. i Hypoxia induces HL-60N cells into VEGFA hi neutrophil-like cells. HL-60N cells were exposed to hypoxia, and the expression levels of VEGFA and ADM were measured by RT–qPCR. j A Venn diagram of the significant transcription factors in SPP1 hi Mφs, VEGFA hi neutrophils and region cluster 1 that were identified by pySCENIC. ATF4 and RUNX3 are activated in all three.

Article Snippet: The primary antibodies were as follows: SPP1 (HPA027541; Atlas; 1:300), ADM (TA806874; Origene; 1:2,000), VEGF (sc-7269; Santa Cruz; 1:4,000); CD68 (ab955; abcam; 1:12,000); CD66b (NB1000-77808; Novus; 1:400), Apelin (ab230536; abcam; 1:50).

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

Journal: Experimental & Molecular Medicine

Article Title: Single-cell transcriptomics of the myeloid milieu reveals an angiogenic niche in triple-negative breast cancer

doi: 10.1038/s12276-025-01571-5

Figure Lengend Snippet: a , b Validation and quantification of our neutrophil subtypes ( a ) and monocyte/macrophage subtypes ( b ) in the TCGA and METABRIC cohorts. The X-bearing boxes indicate the proportion of these subtypes in our scRNA-seq atlas. c Hierarchical clustering of the TNBC patients in the METABRIC cohort on the basis of correlations between the monocyte/macrophage subtype proportions and the clinical metadata. d , e A Kaplan–Meier plot depicting the OS ( d ) and RFS ( e ) of the patients with TNBC in METABRIC after their stratification into the clusters shown in c The cluster-5 patients were enriched in SPP1 hi Mφs and showed worse prognosis than the other patients. By contrast, the cluster-3 patients, who were enriched in CCL3 hi Mφs, had a better prognosis, although this difference did not achieve statistical significance.

Article Snippet: The primary antibodies were as follows: SPP1 (HPA027541; Atlas; 1:300), ADM (TA806874; Origene; 1:2,000), VEGF (sc-7269; Santa Cruz; 1:4,000); CD68 (ab955; abcam; 1:12,000); CD66b (NB1000-77808; Novus; 1:400), Apelin (ab230536; abcam; 1:50).

Techniques: Biomarker Discovery

Journal: Experimental & Molecular Medicine

Article Title: Single-cell transcriptomics of the myeloid milieu reveals an angiogenic niche in triple-negative breast cancer

doi: 10.1038/s12276-025-01571-5

Figure Lengend Snippet: a A schematic depiction of the generation of the EO771 orthotopic breast-cancer model and the downstream scRNA-seq analysis. b UMAP of the monocytes/macrophages in the EO771 tumors. The monocytes/macrophages clustered into three monocyte subtypes, six macrophage subtypes and one DC subtype. c Scoring of the human monocytes/macrophages subtype markers in the EO771 tumors. Only mouse orthologs were used. d Pathway activities of the monocytes/macrophages subtypes in the EO771 tumors, as estimated with GSVA. e UMAP projection of the monocytes/macrophages sorted from the publicly available EMT6 model ( GSE191246 ). The monocytes/macrophages in the EMT6 tumors subclustered into three monocyte subtypes, seven macrophage subtypes and one intermediate subtype. f Hierarchical clustering of the monocyte/macrophage subtypes in human and murine TNBC. The SPP1 hi Mφs are conserved in both humans and mice. g A PCA plot of the pseudobulk data derived from scRNA-seq of EMT6 tumors that had been treated with different anticancer regimens. Group 2 is characterized by C140 treatment. h The proportion of monocytes/macrophages in the different treatment conditions. SPP1 hi Mφs are decreased after C140 treatment. PD1, αPD-1 ICI; Pacli, paclitaxel; Doxo, doxorubicin; Vino, vinorelbine.

Article Snippet: The primary antibodies were as follows: SPP1 (HPA027541; Atlas; 1:300), ADM (TA806874; Origene; 1:2,000), VEGF (sc-7269; Santa Cruz; 1:4,000); CD68 (ab955; abcam; 1:12,000); CD66b (NB1000-77808; Novus; 1:400), Apelin (ab230536; abcam; 1:50).

Techniques: Derivative Assay

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: GSEA results showing the Toll-like receptor signaling pathway is a differentially enriched pathway in the three TNBC. ( A ) KEGG pathway annotations of the Toll-like receptor signaling pathway. ( B ) NES (normalized enrichment score) of each Toll-like receptor signaling pathway-related genes. ( C ) Data for the positive association genes are visualized in a heat map. SPP1 (OPN) is marked with an asterisk.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques:

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: The mRNA expression of SPP1 across various cancer types and their corresponding normal tissues. * p < 0.05, ** p < 0.01, *** p < 0.001. Red boxes denote tumor tissues, while blue boxes represent normal tissues. The red highlights SPP1 expression in tumor tissues, and the blue indicates SPP1 expression in normal tissues. ACC: Adrenocortical carcinoma. BLCA: Bladder urothelial carcinoma. BRCA: Breast invasive carcinoma. CESC: Cervical squamous cell carcinoma and endocervical adenocarcinoma. CHOL: Cholangiocarcinoma. COAD: Colon adenocarcinoma. DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma. ESCA: Esophageal carcinoma. GBM: Glioblastoma multiforme. HNSC: Head and neck squamous cell carcinoma. KICH: Kidney chromophobe. KIRC: Kidney renal clear cell carcinoma. KIRP: Kidney renal papillary cell carcinoma. LAML: Acute myeloid leukemia. LGG: Brain lower-grade glioma. LIHC: Liver hepatocellular carcinoma. LUAD: Lung adenocarcinoma. LUSC: Lung squamous cell carcinoma. MESO: Mesothelioma. OV: Ovarian serous cystadenocarcinoma. PAAD: Pancreatic adenocarcinoma. PCPG: Pheochromocytoma and paraganglioma. PRAD: Prostate adenocarcinoma. READ: Rectum adenocarcinoma. SARC: Sarcoma. SKCM: Skin cutaneous melanoma. STAD: Stomach adenocarcinoma. TGCT: Testicular germ cell tumors. THCA: Thyroid carcinoma. THYM: Thymoma. UCEC: Uterine corpus endometrial carcinoma. UCS: Uterine carcinosarcoma. UVM: Uveal melanoma.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques: Expressing

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: Spearman correlations between the expression of SPP1 and the 32 Toll-like receptor signaling pathway genes in 191 TNBC patients.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques: Expressing

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: Spearman correlation between SPP1 expression and BX795 sensitivity in TNBC cells.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques: Expressing

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: Higher SPP1 mRNA expression had a poor prognosis in BRCA and TNBC. ( A ) Kaplan–Meier survival curve comparing the overall survival (OS) between the low and high SPP1 mRNA expression cohorts in BRCA patients. ( B ) Kaplan–Meier survival curve comparing the relapse-free survival (RFS) between the low and high SPP1 mRNA expression cohorts in BRCA patients. ( C ) Kaplan–Meier survival curve comparing the overall survival (OS) between the low and high SPP1 mRNA expression cohorts in TNBC patients. ( D ) Kaplan–Meier survival curve comparing the relapse-free survival (RFS) between the low and high SPP1 mRNA expression cohorts in TNBC patients.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques: Expressing

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: Immunohistochemical analysis of normal and tumor cell expression of SPP1 from representative samples (100×). Panel shows the number of samples evaluated by the specific SPP1 antibody and that they were positive for SPP1 expression (+) or negative (−). ( A ) The left panel shows representative results of low SPP1 immunostaining in normal breast tissue, while the right panel illustrates high SPP1 immunostaining. ( B ) The left panel displays representative results of low SPP1 immunostaining in breast tumor tissue, while the right panel shows high SPP1 immunostaining. ( C ) SPP1 is not significantly expressed in BRCA tissue compared to its matched normal breast tissue. ( D ) SPP1 is downregulated in TNBC compared to its corresponding normal breast tissues.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques: Immunohistochemical staining, Expressing, Immunostaining

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: SPP1 protein expression had no prognostic significance in BRCA and TNBC. ( A ) Kaplan–Meier plot of the overall survival (OS) was applied to the low and high SPP1 protein cohorts in BRCA patients. ( B ) Kaplan–Meier plot of the relapse-free survival (RFS) was applied to the low and high SPP1 protein cohorts in BRCA patients. ( C ) Kaplan–Meier plot of the overall survival (OS) was applied to the low and high SPP1 protein cohorts in TNBC patients. ( D ) Kaplan–Meier plot of the relapse-free survival (RFS) was applied to the low and high SPP1 protein cohorts in TNBC patients.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques: Expressing

Journal: Current Issues in Molecular Biology

Article Title: SPP1 mRNA Expression Is Associated with M2 Macrophage Infiltration and Poor Prognosis in Triple-Negative Breast Cancer

doi: 10.3390/cimb46120806

Figure Lengend Snippet: Associations between SPP1 mRNA expression and the infiltration of different macrophage subtypes in TNBC: ( A ) M0 macrophages, ( B ) M1 macrophages, and ( C ) M2 macrophages.

Article Snippet: Afterward, the slides were rinsed under gently running tap water for 5 min and then placed in a PBS wash bath for 30 min. Immunohistochemistry (IHC) staining was performed using a primary mouse anti-human SPP1 monoclonal antibody (osteopontin/OPN/SPP1 antibody, AKm2A1: sc-21742; Santa Cruz Biotechnology, Inc. , Dallas, TX, USA) applied at a concentration of 1:200.

Techniques: Expressing