human opn Search Results


recombinant human opn protein  (R&D Systems)
95
R&D Systems recombinant human opn protein
Recombinant Human Opn Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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osteopontin  (R&D Systems)
92
R&D Systems osteopontin
Osteopontin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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elisa kit  (R&D Systems)
94
R&D Systems elisa kit
Fig. 1 hOPN concentrations in milk samples. Human OPN concen- trations in breast milk samples were measured using an <t>ELISA</t> <t>kit</t> (R&D Systems) following the manufacturer’s instructions. Results are shown as means ± SD, n = 10–12 at each time point
Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rh spp1  (R&D Systems)
95
R&D Systems rh spp1
(A) CCL20 protein levels in 786-O RCC cells after induction of EMT by TGFb. (B) DGE between macrophages within 50um of sRCC cells (right) vs >50um (left) from CosMx analysis. (C) Box plot of average <t>SPP1</t> expression of macrophages in Clear Cell, Transition, or Sarcomatoid FOVs from CosMx analysis. (D) Macrophage differentiation marker and SPP1 protein expression in polarized THP-1 derived macrophages and M0 macrophages treated with recombinant CCL20. (E) EMT protein markers from 786-O cells treated with recombinant SPP1. (F) EMT markers in 786-O cells treated with conditioned media from M0, M1, or M2-like macrophages. (G) EMT marker proteins and several proteins found to be highly differentially expressed in Sarcomatoid cells at the mRNA level (CD44, Met, ANXA2) in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (H) PD-L1 protein expression in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (I) Cell invasion assay representative images and quantification in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (J) Schematic of proposed mechanistic pathway of sarcomatoid transformation. Clear cells undergo transformation to “transition” phenotype in which they express increased cytokines including CCL20 which attract immune infiltrate including macrophages. CCL20 and other cytokines lead to M2-like polarization of macrophages with increased SPP1 expression. SPP1+ macrophages then feedback to tumor cells, inducing EMT and PD-L1 expression and leading to full sarcomatoid-like changes.
Rh Spp1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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spp1  (R&D Systems)
93
R&D Systems spp1
Figure 4. High levels of lipid metabolism <t>SPP1+</t> TAM are enriched in metastatic lymph nodes. A) UMAP plot of 3243 TAMs displays the components and relative abundance of cells subtypes, color-coded by subclusters (left) and annotated sample source (right). B) Differential expression of functional state signatures for myeloid cells subclusters and stacked bar plot displays the cell number of myeloid cells with distinct states, color-coded by their sample source. Dot size: percent of cells expressing gene and dot color: scaled average expression. C) Heatmap displays the scaled mean expression of genes associated with functional gene of TAMs D) Heatmap displays the selected signaling pathways (rows) that were significantly enriched in GSVA analyses for each TAMs cluster (columns). Green, blue, and orange purple squares on the left column represent the results derived from KEGG, GO, and REACTOME signaling pathways analysis, respectively. E) Volcano plot displays differentially expressed genes between TAM1_SPP1 (red dots) and the remaining TAMs (blue dots). F) Representative IF staining images displays the expression intensity of SPP1 and CD68 in MLN, NLN and PT (left). Histogram illustrated the fraction of SPP1+ CD68+ TAMs, in matched PT, MLN, and NLN pairs. Data presented as mean ± SEM, N = 5,The P values were calculated by one-way ANOVA test. *p < 0.01, **p < 0.001.
Spp1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human opn elisa kit  (R&D Systems)
95
R&D Systems human opn elisa kit
Figure 4. High levels of lipid metabolism <t>SPP1+</t> TAM are enriched in metastatic lymph nodes. A) UMAP plot of 3243 TAMs displays the components and relative abundance of cells subtypes, color-coded by subclusters (left) and annotated sample source (right). B) Differential expression of functional state signatures for myeloid cells subclusters and stacked bar plot displays the cell number of myeloid cells with distinct states, color-coded by their sample source. Dot size: percent of cells expressing gene and dot color: scaled average expression. C) Heatmap displays the scaled mean expression of genes associated with functional gene of TAMs D) Heatmap displays the selected signaling pathways (rows) that were significantly enriched in GSVA analyses for each TAMs cluster (columns). Green, blue, and orange purple squares on the left column represent the results derived from KEGG, GO, and REACTOME signaling pathways analysis, respectively. E) Volcano plot displays differentially expressed genes between TAM1_SPP1 (red dots) and the remaining TAMs (blue dots). F) Representative IF staining images displays the expression intensity of SPP1 and CD68 in MLN, NLN and PT (left). Histogram illustrated the fraction of SPP1+ CD68+ TAMs, in matched PT, MLN, and NLN pairs. Data presented as mean ± SEM, N = 5,The P values were calculated by one-way ANOVA test. *p < 0.01, **p < 0.001.
Human Opn Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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clone mopc 21  (Bio X Cell)
94
Bio X Cell clone mopc 21
Figure 4. High levels of lipid metabolism <t>SPP1+</t> TAM are enriched in metastatic lymph nodes. A) UMAP plot of 3243 TAMs displays the components and relative abundance of cells subtypes, color-coded by subclusters (left) and annotated sample source (right). B) Differential expression of functional state signatures for myeloid cells subclusters and stacked bar plot displays the cell number of myeloid cells with distinct states, color-coded by their sample source. Dot size: percent of cells expressing gene and dot color: scaled average expression. C) Heatmap displays the scaled mean expression of genes associated with functional gene of TAMs D) Heatmap displays the selected signaling pathways (rows) that were significantly enriched in GSVA analyses for each TAMs cluster (columns). Green, blue, and orange purple squares on the left column represent the results derived from KEGG, GO, and REACTOME signaling pathways analysis, respectively. E) Volcano plot displays differentially expressed genes between TAM1_SPP1 (red dots) and the remaining TAMs (blue dots). F) Representative IF staining images displays the expression intensity of SPP1 and CD68 in MLN, NLN and PT (left). Histogram illustrated the fraction of SPP1+ CD68+ TAMs, in matched PT, MLN, and NLN pairs. Data presented as mean ± SEM, N = 5,The P values were calculated by one-way ANOVA test. *p < 0.01, **p < 0.001.
Clone Mopc 21, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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biotinylated goat anti opn antibody  (R&D Systems)
94
R&D Systems biotinylated goat anti opn antibody
Figure 4. High levels of lipid metabolism <t>SPP1+</t> TAM are enriched in metastatic lymph nodes. A) UMAP plot of 3243 TAMs displays the components and relative abundance of cells subtypes, color-coded by subclusters (left) and annotated sample source (right). B) Differential expression of functional state signatures for myeloid cells subclusters and stacked bar plot displays the cell number of myeloid cells with distinct states, color-coded by their sample source. Dot size: percent of cells expressing gene and dot color: scaled average expression. C) Heatmap displays the scaled mean expression of genes associated with functional gene of TAMs D) Heatmap displays the selected signaling pathways (rows) that were significantly enriched in GSVA analyses for each TAMs cluster (columns). Green, blue, and orange purple squares on the left column represent the results derived from KEGG, GO, and REACTOME signaling pathways analysis, respectively. E) Volcano plot displays differentially expressed genes between TAM1_SPP1 (red dots) and the remaining TAMs (blue dots). F) Representative IF staining images displays the expression intensity of SPP1 and CD68 in MLN, NLN and PT (left). Histogram illustrated the fraction of SPP1+ CD68+ TAMs, in matched PT, MLN, and NLN pairs. Data presented as mean ± SEM, N = 5,The P values were calculated by one-way ANOVA test. *p < 0.01, **p < 0.001.
Biotinylated Goat Anti Opn Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human osteopontin  (R&D Systems)
92
R&D Systems human osteopontin
Fig. 4. A:ThechangesinTGF-b1secretioninprostatecancercelllines,SaOS-2,andbone-derivedstromalcellsbytranilastwereexaminedby ELISA as describedin Materials and Methods Section.Bars, SD.B: <t>Osteopontin</t> (1mg/ml) was addeddaily and 300mmol/L tranilastwas added everyotherdaytoLNCaP-SFcells.Woundphotographsweretakenunderphase-contrastmicroscopyatinitialtime(0hr)andtheterminationof theexperiments(96hr).C:Resultsarepresentedas thepercentagetocontrolin5%CCS(*P < 0.05,**P < 0.01,***P < 0.001).
Human Osteopontin, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse antihuman opn  (R&D Systems)
94
R&D Systems mouse antihuman opn
Fig. 4. A:ThechangesinTGF-b1secretioninprostatecancercelllines,SaOS-2,andbone-derivedstromalcellsbytranilastwereexaminedby ELISA as describedin Materials and Methods Section.Bars, SD.B: <t>Osteopontin</t> (1mg/ml) was addeddaily and 300mmol/L tranilastwas added everyotherdaytoLNCaP-SFcells.Woundphotographsweretakenunderphase-contrastmicroscopyatinitialtime(0hr)andtheterminationof theexperiments(96hr).C:Resultsarepresentedas thepercentagetocontrolin5%CCS(*P < 0.05,**P < 0.01,***P < 0.001).
Mouse Antihuman Opn, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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osteopontin elisa kit  (R&D Systems)
95
R&D Systems osteopontin elisa kit
Fig. 4. A:ThechangesinTGF-b1secretioninprostatecancercelllines,SaOS-2,andbone-derivedstromalcellsbytranilastwereexaminedby ELISA as describedin Materials and Methods Section.Bars, SD.B: <t>Osteopontin</t> (1mg/ml) was addeddaily and 300mmol/L tranilastwas added everyotherdaytoLNCaP-SFcells.Woundphotographsweretakenunderphase-contrastmicroscopyatinitialtime(0hr)andtheterminationof theexperiments(96hr).C:Resultsarepresentedas thepercentagetocontrolin5%CCS(*P < 0.05,**P < 0.01,***P < 0.001).
Osteopontin Elisa Kit, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Fig. 1 hOPN concentrations in milk samples. Human OPN concen- trations in breast milk samples were measured using an ELISA kit (R&D Systems) following the manufacturer’s instructions. Results are shown as means ± SD, n = 10–12 at each time point

Journal: Pediatric research

Article Title: Osteopontin in human milk and infant formula affects infant plasma osteopontin concentrations.

doi: 10.1038/s41390-018-0271-x

Figure Lengend Snippet: Fig. 1 hOPN concentrations in milk samples. Human OPN concen- trations in breast milk samples were measured using an ELISA kit (R&D Systems) following the manufacturer’s instructions. Results are shown as means ± SD, n = 10–12 at each time point

Article Snippet: OPN assays Human OPN (hOPN) concentrations in breast milk and in all infant plasma samples were measured by an ELISA kit (Human Osteopontin DuoSet ELISA, R&D Systems, Minneapolis, MN) following the manufacturer’s instructions. bOPN was also measured in plasma samples by an ELISA kit (Bovine Osteopontin ELISA, LifeSpan BioSciences, Seattle, WA) following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Fig. 2 hOPN concentrations in plasma samples. Human OPN concentrations in all infant plasma samples were measured using a hOPN ELISA kit (R & D Systems) following the manufacturer’s instruction. Plasma samples collected from 1-, 4-, and 6 months old infants (BF, F0, F65, and F130) are shown in a, b, and c, respectively. Results are shown as means ± SD, n = 25 for each treatment group at each time point, *p < 0.001, #p < 0.05, **p < 0.01

Journal: Pediatric research

Article Title: Osteopontin in human milk and infant formula affects infant plasma osteopontin concentrations.

doi: 10.1038/s41390-018-0271-x

Figure Lengend Snippet: Fig. 2 hOPN concentrations in plasma samples. Human OPN concentrations in all infant plasma samples were measured using a hOPN ELISA kit (R & D Systems) following the manufacturer’s instruction. Plasma samples collected from 1-, 4-, and 6 months old infants (BF, F0, F65, and F130) are shown in a, b, and c, respectively. Results are shown as means ± SD, n = 25 for each treatment group at each time point, *p < 0.001, #p < 0.05, **p < 0.01

Article Snippet: OPN assays Human OPN (hOPN) concentrations in breast milk and in all infant plasma samples were measured by an ELISA kit (Human Osteopontin DuoSet ELISA, R&D Systems, Minneapolis, MN) following the manufacturer’s instructions. bOPN was also measured in plasma samples by an ELISA kit (Bovine Osteopontin ELISA, LifeSpan BioSciences, Seattle, WA) following the manufacturer’s instructions.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Fig. 3 bOPN concentrations in plasma samples. The bovine OPN concentration in plasma from 1-, 4-, and 6 months old infants (BF, F0, F65, and F130) was measured using a bovine OPN ELISA kit (Lifespan Biosciences) according to the manufacturer’s instruction. Results are shown means ± SD, n = 8 for each treatment group at each time point. *p < 0.001

Journal: Pediatric research

Article Title: Osteopontin in human milk and infant formula affects infant plasma osteopontin concentrations.

doi: 10.1038/s41390-018-0271-x

Figure Lengend Snippet: Fig. 3 bOPN concentrations in plasma samples. The bovine OPN concentration in plasma from 1-, 4-, and 6 months old infants (BF, F0, F65, and F130) was measured using a bovine OPN ELISA kit (Lifespan Biosciences) according to the manufacturer’s instruction. Results are shown means ± SD, n = 8 for each treatment group at each time point. *p < 0.001

Article Snippet: OPN assays Human OPN (hOPN) concentrations in breast milk and in all infant plasma samples were measured by an ELISA kit (Human Osteopontin DuoSet ELISA, R&D Systems, Minneapolis, MN) following the manufacturer’s instructions. bOPN was also measured in plasma samples by an ELISA kit (Bovine Osteopontin ELISA, LifeSpan BioSciences, Seattle, WA) following the manufacturer’s instructions.

Techniques: Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay

(A) CCL20 protein levels in 786-O RCC cells after induction of EMT by TGFb. (B) DGE between macrophages within 50um of sRCC cells (right) vs >50um (left) from CosMx analysis. (C) Box plot of average SPP1 expression of macrophages in Clear Cell, Transition, or Sarcomatoid FOVs from CosMx analysis. (D) Macrophage differentiation marker and SPP1 protein expression in polarized THP-1 derived macrophages and M0 macrophages treated with recombinant CCL20. (E) EMT protein markers from 786-O cells treated with recombinant SPP1. (F) EMT markers in 786-O cells treated with conditioned media from M0, M1, or M2-like macrophages. (G) EMT marker proteins and several proteins found to be highly differentially expressed in Sarcomatoid cells at the mRNA level (CD44, Met, ANXA2) in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (H) PD-L1 protein expression in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (I) Cell invasion assay representative images and quantification in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (J) Schematic of proposed mechanistic pathway of sarcomatoid transformation. Clear cells undergo transformation to “transition” phenotype in which they express increased cytokines including CCL20 which attract immune infiltrate including macrophages. CCL20 and other cytokines lead to M2-like polarization of macrophages with increased SPP1 expression. SPP1+ macrophages then feedback to tumor cells, inducing EMT and PD-L1 expression and leading to full sarcomatoid-like changes.

Journal: bioRxiv

Article Title: Spatial analysis reveals a novel inflammatory tumor transition state which promotes a macrophage-driven induction of sarcomatoid renal cell carcinoma

doi: 10.1101/2025.06.12.656421

Figure Lengend Snippet: (A) CCL20 protein levels in 786-O RCC cells after induction of EMT by TGFb. (B) DGE between macrophages within 50um of sRCC cells (right) vs >50um (left) from CosMx analysis. (C) Box plot of average SPP1 expression of macrophages in Clear Cell, Transition, or Sarcomatoid FOVs from CosMx analysis. (D) Macrophage differentiation marker and SPP1 protein expression in polarized THP-1 derived macrophages and M0 macrophages treated with recombinant CCL20. (E) EMT protein markers from 786-O cells treated with recombinant SPP1. (F) EMT markers in 786-O cells treated with conditioned media from M0, M1, or M2-like macrophages. (G) EMT marker proteins and several proteins found to be highly differentially expressed in Sarcomatoid cells at the mRNA level (CD44, Met, ANXA2) in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (H) PD-L1 protein expression in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (I) Cell invasion assay representative images and quantification in 786-O cells treated with conditioned media from M2 macrophages with or without SPP1 knockdown by siRNA. (J) Schematic of proposed mechanistic pathway of sarcomatoid transformation. Clear cells undergo transformation to “transition” phenotype in which they express increased cytokines including CCL20 which attract immune infiltrate including macrophages. CCL20 and other cytokines lead to M2-like polarization of macrophages with increased SPP1 expression. SPP1+ macrophages then feedback to tumor cells, inducing EMT and PD-L1 expression and leading to full sarcomatoid-like changes.

Article Snippet: We purchased Recombinant Human (rh) TGF-beta 1 (7754-BH), rh SPP1 (1433-OP), and rh CCL20 (360-MP) from R&D systems, rh Interleukin-4 (IL-4) (ab155733) from Abcam, lipopolysaccharide (LPS) (L4391) from Sigma-Aldrich, and rh Interferon (IFN)-γ (PHC4031) from Thermo Fisher Scientific.

Techniques: Expressing, Marker, Derivative Assay, Recombinant, Knockdown, Invasion Assay, Transformation Assay

Western blot showing EMT markers in UOK-127 cells treated with 50ng/mL recombinant SPP1 at varying timepoints.

Journal: bioRxiv

Article Title: Spatial analysis reveals a novel inflammatory tumor transition state which promotes a macrophage-driven induction of sarcomatoid renal cell carcinoma

doi: 10.1101/2025.06.12.656421

Figure Lengend Snippet: Western blot showing EMT markers in UOK-127 cells treated with 50ng/mL recombinant SPP1 at varying timepoints.

Article Snippet: We purchased Recombinant Human (rh) TGF-beta 1 (7754-BH), rh SPP1 (1433-OP), and rh CCL20 (360-MP) from R&D systems, rh Interleukin-4 (IL-4) (ab155733) from Abcam, lipopolysaccharide (LPS) (L4391) from Sigma-Aldrich, and rh Interferon (IFN)-γ (PHC4031) from Thermo Fisher Scientific.

Techniques: Western Blot, Recombinant

Western blot showing SPP1 expression in THP-1 derived M2-like macrophages including control, and treatment with silencing mRNAs siNC, siPC, siSPP1#1, and siSPP1#2.

Journal: bioRxiv

Article Title: Spatial analysis reveals a novel inflammatory tumor transition state which promotes a macrophage-driven induction of sarcomatoid renal cell carcinoma

doi: 10.1101/2025.06.12.656421

Figure Lengend Snippet: Western blot showing SPP1 expression in THP-1 derived M2-like macrophages including control, and treatment with silencing mRNAs siNC, siPC, siSPP1#1, and siSPP1#2.

Article Snippet: We purchased Recombinant Human (rh) TGF-beta 1 (7754-BH), rh SPP1 (1433-OP), and rh CCL20 (360-MP) from R&D systems, rh Interleukin-4 (IL-4) (ab155733) from Abcam, lipopolysaccharide (LPS) (L4391) from Sigma-Aldrich, and rh Interferon (IFN)-γ (PHC4031) from Thermo Fisher Scientific.

Techniques: Western Blot, Expressing, Derivative Assay, Control

Western blot showing sarcomatoid-specific proteins (Met, CD44, ANXA2) and EMT markers (N-cadherin, Slug, Snail) in UOK-127 cells control, treated with M2-like macrophages, or treated with M2-like macrophages with siRNA knockdown of negative control or SPP1.

Journal: bioRxiv

Article Title: Spatial analysis reveals a novel inflammatory tumor transition state which promotes a macrophage-driven induction of sarcomatoid renal cell carcinoma

doi: 10.1101/2025.06.12.656421

Figure Lengend Snippet: Western blot showing sarcomatoid-specific proteins (Met, CD44, ANXA2) and EMT markers (N-cadherin, Slug, Snail) in UOK-127 cells control, treated with M2-like macrophages, or treated with M2-like macrophages with siRNA knockdown of negative control or SPP1.

Article Snippet: We purchased Recombinant Human (rh) TGF-beta 1 (7754-BH), rh SPP1 (1433-OP), and rh CCL20 (360-MP) from R&D systems, rh Interleukin-4 (IL-4) (ab155733) from Abcam, lipopolysaccharide (LPS) (L4391) from Sigma-Aldrich, and rh Interferon (IFN)-γ (PHC4031) from Thermo Fisher Scientific.

Techniques: Western Blot, Control, Knockdown, Negative Control

Invasion assay of UOK-127 cell co-cultured with M2 macrophage conditioned media with negative controle (siNC) or silencing of SPP1 (siSPP1) in M2 macrophages.

Journal: bioRxiv

Article Title: Spatial analysis reveals a novel inflammatory tumor transition state which promotes a macrophage-driven induction of sarcomatoid renal cell carcinoma

doi: 10.1101/2025.06.12.656421

Figure Lengend Snippet: Invasion assay of UOK-127 cell co-cultured with M2 macrophage conditioned media with negative controle (siNC) or silencing of SPP1 (siSPP1) in M2 macrophages.

Article Snippet: We purchased Recombinant Human (rh) TGF-beta 1 (7754-BH), rh SPP1 (1433-OP), and rh CCL20 (360-MP) from R&D systems, rh Interleukin-4 (IL-4) (ab155733) from Abcam, lipopolysaccharide (LPS) (L4391) from Sigma-Aldrich, and rh Interferon (IFN)-γ (PHC4031) from Thermo Fisher Scientific.

Techniques: Invasion Assay, Cell Culture

Figure 4. High levels of lipid metabolism SPP1+ TAM are enriched in metastatic lymph nodes. A) UMAP plot of 3243 TAMs displays the components and relative abundance of cells subtypes, color-coded by subclusters (left) and annotated sample source (right). B) Differential expression of functional state signatures for myeloid cells subclusters and stacked bar plot displays the cell number of myeloid cells with distinct states, color-coded by their sample source. Dot size: percent of cells expressing gene and dot color: scaled average expression. C) Heatmap displays the scaled mean expression of genes associated with functional gene of TAMs D) Heatmap displays the selected signaling pathways (rows) that were significantly enriched in GSVA analyses for each TAMs cluster (columns). Green, blue, and orange purple squares on the left column represent the results derived from KEGG, GO, and REACTOME signaling pathways analysis, respectively. E) Volcano plot displays differentially expressed genes between TAM1_SPP1 (red dots) and the remaining TAMs (blue dots). F) Representative IF staining images displays the expression intensity of SPP1 and CD68 in MLN, NLN and PT (left). Histogram illustrated the fraction of SPP1+ CD68+ TAMs, in matched PT, MLN, and NLN pairs. Data presented as mean ± SEM, N = 5,The P values were calculated by one-way ANOVA test. *p < 0.01, **p < 0.001.

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: SPP1 + TAM Regulates the Metastatic Colonization of CXCR4 + Metastasis-Associated Tumor Cells by Remodeling the Lymph Node Microenvironment.

doi: 10.1002/advs.202400524

Figure Lengend Snippet: Figure 4. High levels of lipid metabolism SPP1+ TAM are enriched in metastatic lymph nodes. A) UMAP plot of 3243 TAMs displays the components and relative abundance of cells subtypes, color-coded by subclusters (left) and annotated sample source (right). B) Differential expression of functional state signatures for myeloid cells subclusters and stacked bar plot displays the cell number of myeloid cells with distinct states, color-coded by their sample source. Dot size: percent of cells expressing gene and dot color: scaled average expression. C) Heatmap displays the scaled mean expression of genes associated with functional gene of TAMs D) Heatmap displays the selected signaling pathways (rows) that were significantly enriched in GSVA analyses for each TAMs cluster (columns). Green, blue, and orange purple squares on the left column represent the results derived from KEGG, GO, and REACTOME signaling pathways analysis, respectively. E) Volcano plot displays differentially expressed genes between TAM1_SPP1 (red dots) and the remaining TAMs (blue dots). F) Representative IF staining images displays the expression intensity of SPP1 and CD68 in MLN, NLN and PT (left). Histogram illustrated the fraction of SPP1+ CD68+ TAMs, in matched PT, MLN, and NLN pairs. Data presented as mean ± SEM, N = 5,The P values were calculated by one-way ANOVA test. *p < 0.01, **p < 0.001.

Article Snippet: The primary antibodies were: CD8 (Cat. 85336S, CST), SPP1 (Cat.AF1433, R&D Systems), CD68 (Cat. ab213363), FOXP3 (Cat. ab20034, Abcam) and GZMB (Cat. ab255598, Abcam).

Techniques: Expressing, Functional Assay, Protein-Protein interactions, Derivative Assay, Staining

Figure 5. SPP1+ TAMs are derived from VCAN+ inflammatory monocytes. A) Pseudotime-ordered analysis of five TAMs by Monocle. B) Pseudotime- ordered analysis marked by TAM1_SPP1 and TAM3_VCAN (Left). CytoTRACE score in TAM3 versus TAM1 (Right). C) 2D pseudotime plot displays the pseudotime score identified by Monocle. D) 2D pseudotime plot displays the CytoTRACE score identified by CytoTRACE. E) Histogram displays the cell distribution of TAMs. TAMs subtypes labeled by colors. F) Heatmap displays the dynamic changes in gene expression along the pseudotime (lower panel). The distribution of TAMs subtypes during the transition (divided into four phases), along with the pseudo-time. Subtypes were labeled by colors (upper panel). 2D plots showed the loess regression-smoothened dynamic expression (y-axis) of transcription factor G), histocompatibility complexes or immunosuppressive molecules H), and lipid metabolism genes I) during the TAMs transitions along the pseudotime. Green, TAM1_SPP1; blue, TAM3_VCAN.

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: SPP1 + TAM Regulates the Metastatic Colonization of CXCR4 + Metastasis-Associated Tumor Cells by Remodeling the Lymph Node Microenvironment.

doi: 10.1002/advs.202400524

Figure Lengend Snippet: Figure 5. SPP1+ TAMs are derived from VCAN+ inflammatory monocytes. A) Pseudotime-ordered analysis of five TAMs by Monocle. B) Pseudotime- ordered analysis marked by TAM1_SPP1 and TAM3_VCAN (Left). CytoTRACE score in TAM3 versus TAM1 (Right). C) 2D pseudotime plot displays the pseudotime score identified by Monocle. D) 2D pseudotime plot displays the CytoTRACE score identified by CytoTRACE. E) Histogram displays the cell distribution of TAMs. TAMs subtypes labeled by colors. F) Heatmap displays the dynamic changes in gene expression along the pseudotime (lower panel). The distribution of TAMs subtypes during the transition (divided into four phases), along with the pseudo-time. Subtypes were labeled by colors (upper panel). 2D plots showed the loess regression-smoothened dynamic expression (y-axis) of transcription factor G), histocompatibility complexes or immunosuppressive molecules H), and lipid metabolism genes I) during the TAMs transitions along the pseudotime. Green, TAM1_SPP1; blue, TAM3_VCAN.

Article Snippet: The primary antibodies were: CD8 (Cat. 85336S, CST), SPP1 (Cat.AF1433, R&D Systems), CD68 (Cat. ab213363), FOXP3 (Cat. ab20034, Abcam) and GZMB (Cat. ab255598, Abcam).

Techniques: Derivative Assay, Labeling, Gene Expression, Expressing

Figure 6. Different tumor immune phenotypes between primary tumor and metastatic lymph node shaped by SPP1+ TAM. A) Representative images of mIHC staining for the Treg cells (FOXP3+), Cytotoxic T cells (CD8+ GZMB+), and SPP1+ CD68+ TAMs in PT and MLN. Images are representative of ten biological replicates. B) Top, the fraction of SPP1+ CD68+ TAMs in both tumor nest, and stroma between PT and MLN (N = 14). Bottom, the fold change of FOXP3+ Treg to GZMB+ CD8+ T cells ratio in both tumor nest, and stroma between PT and MLN (N = 14). Data presented as mean ± SEM. The P values were calculated by one-way ANOVA test. ns, not significant, **p < 0.001, ****p < 0.00001. C) Dot plots displays selected ligand-receptor interactions between TAM1_SPP1 and T cells, as well as NK cells. The ligand-receptor interactions and cell-cell interactions are indicated at columns and

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: SPP1 + TAM Regulates the Metastatic Colonization of CXCR4 + Metastasis-Associated Tumor Cells by Remodeling the Lymph Node Microenvironment.

doi: 10.1002/advs.202400524

Figure Lengend Snippet: Figure 6. Different tumor immune phenotypes between primary tumor and metastatic lymph node shaped by SPP1+ TAM. A) Representative images of mIHC staining for the Treg cells (FOXP3+), Cytotoxic T cells (CD8+ GZMB+), and SPP1+ CD68+ TAMs in PT and MLN. Images are representative of ten biological replicates. B) Top, the fraction of SPP1+ CD68+ TAMs in both tumor nest, and stroma between PT and MLN (N = 14). Bottom, the fold change of FOXP3+ Treg to GZMB+ CD8+ T cells ratio in both tumor nest, and stroma between PT and MLN (N = 14). Data presented as mean ± SEM. The P values were calculated by one-way ANOVA test. ns, not significant, **p < 0.001, ****p < 0.00001. C) Dot plots displays selected ligand-receptor interactions between TAM1_SPP1 and T cells, as well as NK cells. The ligand-receptor interactions and cell-cell interactions are indicated at columns and

Article Snippet: The primary antibodies were: CD8 (Cat. 85336S, CST), SPP1 (Cat.AF1433, R&D Systems), CD68 (Cat. ab213363), FOXP3 (Cat. ab20034, Abcam) and GZMB (Cat. ab255598, Abcam).

Techniques: Staining

Figure 7. Heterogeneity of macrophages across various metastatic sites in different types of cancer. A) UMAP plot illustrates the distribution of macrophages in various tumor metastases, color-coded by subclusters. B) The bar plot displays the proportion of macrophage subpopulations across different tumor metastasis sites. C) The selected genes expression in TAM1_SPP1 from different sample sources. Each column in the heatmap repre- sents a patient. Labels above the heatmap indicates the sample source and tumor type. D) Venn diagram illustrates the intersection of highly expressed genes in APOC1+ APOE+ macrophages in ESCC and TAM1_SPP1 in OSCC. The specific 14 intersection genes are listed below. E) The violin plot displays the expression scores of APOC1+ APOE+ macrophages in TAM subpopulations in our manuscript. Box plots inside the violins indicate the quartiles of corresponding score levels.

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: SPP1 + TAM Regulates the Metastatic Colonization of CXCR4 + Metastasis-Associated Tumor Cells by Remodeling the Lymph Node Microenvironment.

doi: 10.1002/advs.202400524

Figure Lengend Snippet: Figure 7. Heterogeneity of macrophages across various metastatic sites in different types of cancer. A) UMAP plot illustrates the distribution of macrophages in various tumor metastases, color-coded by subclusters. B) The bar plot displays the proportion of macrophage subpopulations across different tumor metastasis sites. C) The selected genes expression in TAM1_SPP1 from different sample sources. Each column in the heatmap repre- sents a patient. Labels above the heatmap indicates the sample source and tumor type. D) Venn diagram illustrates the intersection of highly expressed genes in APOC1+ APOE+ macrophages in ESCC and TAM1_SPP1 in OSCC. The specific 14 intersection genes are listed below. E) The violin plot displays the expression scores of APOC1+ APOE+ macrophages in TAM subpopulations in our manuscript. Box plots inside the violins indicate the quartiles of corresponding score levels.

Article Snippet: The primary antibodies were: CD8 (Cat. 85336S, CST), SPP1 (Cat.AF1433, R&D Systems), CD68 (Cat. ab213363), FOXP3 (Cat. ab20034, Abcam) and GZMB (Cat. ab255598, Abcam).

Techniques: Expressing

Fig. 4. A:ThechangesinTGF-b1secretioninprostatecancercelllines,SaOS-2,andbone-derivedstromalcellsbytranilastwereexaminedby ELISA as describedin Materials and Methods Section.Bars, SD.B: Osteopontin (1mg/ml) was addeddaily and 300mmol/L tranilastwas added everyotherdaytoLNCaP-SFcells.Woundphotographsweretakenunderphase-contrastmicroscopyatinitialtime(0hr)andtheterminationof theexperiments(96hr).C:Resultsarepresentedas thepercentagetocontrolin5%CCS(*P < 0.05,**P < 0.01,***P < 0.001).

Journal: The Prostate

Article Title: Tranilast inhibits hormone refractory prostate cancer cell proliferation and suppresses transforming growth factor beta1-associated osteoblastic changes.

doi: 10.1002/pros.20975

Figure Lengend Snippet: Fig. 4. A:ThechangesinTGF-b1secretioninprostatecancercelllines,SaOS-2,andbone-derivedstromalcellsbytranilastwereexaminedby ELISA as describedin Materials and Methods Section.Bars, SD.B: Osteopontin (1mg/ml) was addeddaily and 300mmol/L tranilastwas added everyotherdaytoLNCaP-SFcells.Woundphotographsweretakenunderphase-contrastmicroscopyatinitialtime(0hr)andtheterminationof theexperiments(96hr).C:Resultsarepresentedas thepercentagetocontrolin5%CCS(*P < 0.05,**P < 0.01,***P < 0.001).

Article Snippet: Cells were treated with recombinant human osteopontin (R&D Systems) alone or with tranilast in 5% CCS or serum free medium.

Techniques: Enzyme-linked Immunosorbent Assay