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    ATCC l wnt 3a
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    l wnt 3a cells  (ATCC)


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    ATCC l wnt 3a cells
    Human monocytes <t>express</t> <t>Wnt</t> signaling components and the <t>Wnt-3a</t> ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve
    L Wnt 3a Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Wnt signaling regulates chemokine production and cell migration of circulating human monocytes"

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-024-01608-8

    Human monocytes express Wnt signaling components and the Wnt-3a ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve
    Figure Legend Snippet: Human monocytes express Wnt signaling components and the Wnt-3a ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve

    Techniques Used: RNA Sequencing Assay, Labeling, Expressing, Functional Assay, Enzyme-linked Immunosorbent Assay, Isolation

    Wnt-3a induces expression and nuclear accumulation of β-catenin in monocytes. A and B , THP-1 cells (monocyte-like cells), PMA-treated THP-1 cells (macrophage-like cells), and freshly isolated primary monocytes were cultured for 5 h in the presence of control or Wnt-3a conditioned media (CM). A Cells were lysed, and western blot analysis was conducted using specific antibodies as indicated (representative blot). B Quantification of band intensity of experiments as in A, calculated by the ImageJ software. The Y-axis represents the band intensity of β-catenin standardized to actin (β-catenin/actin) of Wnt-treated cells relative to control-treated cells (100%). C-E Treated monocytes (as in A) were stained and visualized using immunofluorescence. Images were obtained using a confocal microscope with a 60X oil-immersion objective lens C . An independent script was used to quantify the signal intensity and nuclear localization of β-catenin. Pairs signify individual donors, where each point represents the mean value of at least 5 fields D, E (see methods section). * P -value = 0.0359 for paired t-test of signal intensity D and * P -value = 0.0342 for paired t-test of nuclear localization E
    Figure Legend Snippet: Wnt-3a induces expression and nuclear accumulation of β-catenin in monocytes. A and B , THP-1 cells (monocyte-like cells), PMA-treated THP-1 cells (macrophage-like cells), and freshly isolated primary monocytes were cultured for 5 h in the presence of control or Wnt-3a conditioned media (CM). A Cells were lysed, and western blot analysis was conducted using specific antibodies as indicated (representative blot). B Quantification of band intensity of experiments as in A, calculated by the ImageJ software. The Y-axis represents the band intensity of β-catenin standardized to actin (β-catenin/actin) of Wnt-treated cells relative to control-treated cells (100%). C-E Treated monocytes (as in A) were stained and visualized using immunofluorescence. Images were obtained using a confocal microscope with a 60X oil-immersion objective lens C . An independent script was used to quantify the signal intensity and nuclear localization of β-catenin. Pairs signify individual donors, where each point represents the mean value of at least 5 fields D, E (see methods section). * P -value = 0.0359 for paired t-test of signal intensity D and * P -value = 0.0342 for paired t-test of nuclear localization E

    Techniques Used: Expressing, Isolation, Cell Culture, Western Blot, Software, Staining, Immunofluorescence, Microscopy

    Wnt-3a does not induce expression of classical Wnt signaling target genes in monocytes. A Treated monocytes (as in Fig. ) were lysed for RT-qPCR analysis of the indicated Wnt target genes. B Cell protein lysates were analyzed by western blot using specific antibodies as indicated. The graphs represent the relative band intensity. C Treated monocytes were stained and visualized using immunofluorescence with the indicated antibodies. Signal intensity was quantified with no significant differences for any paired t-test
    Figure Legend Snippet: Wnt-3a does not induce expression of classical Wnt signaling target genes in monocytes. A Treated monocytes (as in Fig. ) were lysed for RT-qPCR analysis of the indicated Wnt target genes. B Cell protein lysates were analyzed by western blot using specific antibodies as indicated. The graphs represent the relative band intensity. C Treated monocytes were stained and visualized using immunofluorescence with the indicated antibodies. Signal intensity was quantified with no significant differences for any paired t-test

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Staining, Immunofluorescence

    Wnt-3a modifies the levels of monocyte-secreted immune proteins. Freshly isolated monocytes (from two donors) were cultured (in triplicates) for 8 h in the presence of control or Wnt-3a conditioned media (CM) for RNA sequencing. A Principal component analysis based on entire gene expression patterns, with batch effect removal to standardize the different donors. B Volcano plot of all genes based on differential expression analysis between control- and Wnt-treated monocytes. Significantly regulated genes are in red on either side (as defined by adjusted p -value < 0.05 and absolute fold change > 2). C, D Summary of Log2 Fold Change of significantly regulated cytokine and chemokine ligands C and receptors D . E Similarly treated monocytes from multiple blood samples were lysed for RT-qPCR assays of the indicated cytokines and chemokines
    Figure Legend Snippet: Wnt-3a modifies the levels of monocyte-secreted immune proteins. Freshly isolated monocytes (from two donors) were cultured (in triplicates) for 8 h in the presence of control or Wnt-3a conditioned media (CM) for RNA sequencing. A Principal component analysis based on entire gene expression patterns, with batch effect removal to standardize the different donors. B Volcano plot of all genes based on differential expression analysis between control- and Wnt-treated monocytes. Significantly regulated genes are in red on either side (as defined by adjusted p -value < 0.05 and absolute fold change > 2). C, D Summary of Log2 Fold Change of significantly regulated cytokine and chemokine ligands C and receptors D . E Similarly treated monocytes from multiple blood samples were lysed for RT-qPCR assays of the indicated cytokines and chemokines

    Techniques Used: Isolation, Cell Culture, RNA Sequencing Assay, Expressing, Quantitative RT-PCR

    Wnt-3a affects chemokine secretion from monocytes. Media collected from the treated monocytes from two donors were centrifuged twice to remove the cells, and subjected to a chemokine membrane array (see methods section). Analysis of a representative membrane is shown in the two left panels. A summary of the changes in the chemokines detected in the two experiments is presented on the right: upregulated (red), downregulated (blue), and unchanged (grey) proteins. Original images and analyses are presented in Fig. S
    Figure Legend Snippet: Wnt-3a affects chemokine secretion from monocytes. Media collected from the treated monocytes from two donors were centrifuged twice to remove the cells, and subjected to a chemokine membrane array (see methods section). Analysis of a representative membrane is shown in the two left panels. A summary of the changes in the chemokines detected in the two experiments is presented on the right: upregulated (red), downregulated (blue), and unchanged (grey) proteins. Original images and analyses are presented in Fig. S

    Techniques Used: Membrane

    Treating monocytes with Wnt-3a induces cell migration and CCL2 production. Naïve monocytes were plated in the top chamber of a transwell and exposed to different conditioned media (CM). Cells that crossed the transwell were collected from the bottom well and counted by flow cytometry. A Schematic illustration of experimental design. Naïve monocytes were exposed to Wnt-3a/control CM obtained from L-cells for 2 h (I). Naïve monocytes were exposed to Wnt-3a/control CM obtained from monocytes treated with Wnt-3a/control for 8 h (II). Treated monocytes (as in II) were washed and transferred to fresh RPMI medium for an additional 1 h. CM was collected and used to treat naïve monocytes in the transwell assay (III). B Flow cytometry counts of cells crossing the transwell. * P -value = 0.0307, *** P -value = 0.0003 and ** P -value = 0.0045 for paired t-tests. Y-axis represents the absolute cell count. C ELISA assays of CCL2 concentration in media used for the indicated experiments. Protein concentrations were calculated by regression from a standard curve. D Flow cytometry counts of cells crossing the transwell. Y-axis represents the absolute cell count. X-axis represents the media used as in I and II of the experimental design, with or without 2 μg/ml CCL2 inhibitor, marked as circles and squares, respectively. The white and black shapes represent each donor. FC – average fold change of the marked pairs. E A representative blot of media collected from the top and bottom chambers of the different experiments (as indicated), at the end of the transwell incubation period. Media were centrifuged to remove cells and analyzed by western blot using a specific anti-Wnt-3a antibody. C – control/control-treated media, W3 – Wnt-3a/Wnt-3a-treated media
    Figure Legend Snippet: Treating monocytes with Wnt-3a induces cell migration and CCL2 production. Naïve monocytes were plated in the top chamber of a transwell and exposed to different conditioned media (CM). Cells that crossed the transwell were collected from the bottom well and counted by flow cytometry. A Schematic illustration of experimental design. Naïve monocytes were exposed to Wnt-3a/control CM obtained from L-cells for 2 h (I). Naïve monocytes were exposed to Wnt-3a/control CM obtained from monocytes treated with Wnt-3a/control for 8 h (II). Treated monocytes (as in II) were washed and transferred to fresh RPMI medium for an additional 1 h. CM was collected and used to treat naïve monocytes in the transwell assay (III). B Flow cytometry counts of cells crossing the transwell. * P -value = 0.0307, *** P -value = 0.0003 and ** P -value = 0.0045 for paired t-tests. Y-axis represents the absolute cell count. C ELISA assays of CCL2 concentration in media used for the indicated experiments. Protein concentrations were calculated by regression from a standard curve. D Flow cytometry counts of cells crossing the transwell. Y-axis represents the absolute cell count. X-axis represents the media used as in I and II of the experimental design, with or without 2 μg/ml CCL2 inhibitor, marked as circles and squares, respectively. The white and black shapes represent each donor. FC – average fold change of the marked pairs. E A representative blot of media collected from the top and bottom chambers of the different experiments (as indicated), at the end of the transwell incubation period. Media were centrifuged to remove cells and analyzed by western blot using a specific anti-Wnt-3a antibody. C – control/control-treated media, W3 – Wnt-3a/Wnt-3a-treated media

    Techniques Used: Migration, Flow Cytometry, Transwell Assay, Cell Counting, Enzyme-linked Immunosorbent Assay, Concentration Assay, Incubation, Western Blot

    The inflammatory state of patients with rheumatic joint diseases affects monocyte reaction to Wnt-3a stimulus. A Principal component analyses (PCA) based on RT-qPCR data (presented in Fig. S B) of Wnt-3a-treated monocytes (treated as in Fig. E). The blue X represents the centroid of the healthy donor samples, calculated as the average of the x and y coordinates of all the blue dots on the PCA graph. The bar graphs represent a calculation of distance of each dot from the healthy donor centroid, based on x,y coordinates. One-way ANOVA with Dunnett's multiple comparisons test to healthy control were employed. Healthy donors (blue), treated RJD patients (pink) and RJD patients naïve to treatment (maroon). ** P -value = 0.0024, ns = 0.6717. B Media were collected from the treated monocytes of two RJD patients naïve to immunosuppressive treatment. The media were subjected to a chemokine membrane array and analyzed, as performed with media of healthy donors in Fig. . The heatmap was created using numerical data of both the data of healthy donors represented in Fig. (Healthy 1&2) and the data of the two RJD patients (RJD 1&2). The fold change (FC) of signal ratio of Wnt-treated to control-treated monocyte is shown as a heatmap (log2(FC)), where upregulated genes are represented in red, downregulated in blue, and unchanged in white. Hierarchical clustering was performed on both the donors on the X-axis and the secreted proteins on the Y-axis. Only proteins clearly detected across all four membranes are represented. Original images and analyses of all 4 donors are presented in Fig. S , Fig. S , and the raw figure data (Supplementary file ). RJD – rheumatic joint diseases
    Figure Legend Snippet: The inflammatory state of patients with rheumatic joint diseases affects monocyte reaction to Wnt-3a stimulus. A Principal component analyses (PCA) based on RT-qPCR data (presented in Fig. S B) of Wnt-3a-treated monocytes (treated as in Fig. E). The blue X represents the centroid of the healthy donor samples, calculated as the average of the x and y coordinates of all the blue dots on the PCA graph. The bar graphs represent a calculation of distance of each dot from the healthy donor centroid, based on x,y coordinates. One-way ANOVA with Dunnett's multiple comparisons test to healthy control were employed. Healthy donors (blue), treated RJD patients (pink) and RJD patients naïve to treatment (maroon). ** P -value = 0.0024, ns = 0.6717. B Media were collected from the treated monocytes of two RJD patients naïve to immunosuppressive treatment. The media were subjected to a chemokine membrane array and analyzed, as performed with media of healthy donors in Fig. . The heatmap was created using numerical data of both the data of healthy donors represented in Fig. (Healthy 1&2) and the data of the two RJD patients (RJD 1&2). The fold change (FC) of signal ratio of Wnt-treated to control-treated monocyte is shown as a heatmap (log2(FC)), where upregulated genes are represented in red, downregulated in blue, and unchanged in white. Hierarchical clustering was performed on both the donors on the X-axis and the secreted proteins on the Y-axis. Only proteins clearly detected across all four membranes are represented. Original images and analyses of all 4 donors are presented in Fig. S , Fig. S , and the raw figure data (Supplementary file ). RJD – rheumatic joint diseases

    Techniques Used: Quantitative RT-PCR, Membrane

    l wnt 3a cells  (ATCC)


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    ATCC l wnt 3a cells
    Human monocytes <t>express</t> <t>Wnt</t> signaling components and the <t>Wnt-3a</t> ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve
    L Wnt 3a Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC cell lines l wnt 3a cell line
    Human monocytes <t>express</t> <t>Wnt</t> signaling components and the <t>Wnt-3a</t> ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve
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    Human monocytes <t>express</t> <t>Wnt</t> signaling components and the <t>Wnt-3a</t> ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve
    L Wnt 3a Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC l wnt 3a conditioned media
    Human monocytes <t>express</t> <t>Wnt</t> signaling components and the <t>Wnt-3a</t> ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve
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    Human monocytes express Wnt signaling components and the Wnt-3a ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve

    Journal: Cell Communication and Signaling : CCS

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    doi: 10.1186/s12964-024-01608-8

    Figure Lengend Snippet: Human monocytes express Wnt signaling components and the Wnt-3a ligand is found in the plasma. A Analysis of single-cell RNA-seq data by 10X genomics, based on a Seurat tutorial (“pbmc3k”). tSNE plot of the data with annotation of cell types (upper left panel). Classical monocytes are the orange cluster labeled “CD14 + Mono” and non-classical monocytes are the light blue cluster labeled “CD16 + Mono”. Plt – platelets. NK – natural killer cells. B and T lymphocytes are abbreviated B and T, respectively. The monocyte cluster (red circle) was further analyzed for expression of Wnt signaling components. The genes are grouped by functional categories (color-coded at the bottom). B Wnt-3a levels were measured by ELISA in isolated human plasma obtained from multiple healthy donors. Protein concentrations were calculated by regression from a standard curve

    Article Snippet: L-Wnt-3a cells, L cells, and THP-1 cells were propagated according to the ATCC guidelines.

    Techniques: RNA Sequencing Assay, Labeling, Expressing, Functional Assay, Enzyme-linked Immunosorbent Assay, Isolation

    Wnt-3a induces expression and nuclear accumulation of β-catenin in monocytes. A and B , THP-1 cells (monocyte-like cells), PMA-treated THP-1 cells (macrophage-like cells), and freshly isolated primary monocytes were cultured for 5 h in the presence of control or Wnt-3a conditioned media (CM). A Cells were lysed, and western blot analysis was conducted using specific antibodies as indicated (representative blot). B Quantification of band intensity of experiments as in A, calculated by the ImageJ software. The Y-axis represents the band intensity of β-catenin standardized to actin (β-catenin/actin) of Wnt-treated cells relative to control-treated cells (100%). C-E Treated monocytes (as in A) were stained and visualized using immunofluorescence. Images were obtained using a confocal microscope with a 60X oil-immersion objective lens C . An independent script was used to quantify the signal intensity and nuclear localization of β-catenin. Pairs signify individual donors, where each point represents the mean value of at least 5 fields D, E (see methods section). * P -value = 0.0359 for paired t-test of signal intensity D and * P -value = 0.0342 for paired t-test of nuclear localization E

    Journal: Cell Communication and Signaling : CCS

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    doi: 10.1186/s12964-024-01608-8

    Figure Lengend Snippet: Wnt-3a induces expression and nuclear accumulation of β-catenin in monocytes. A and B , THP-1 cells (monocyte-like cells), PMA-treated THP-1 cells (macrophage-like cells), and freshly isolated primary monocytes were cultured for 5 h in the presence of control or Wnt-3a conditioned media (CM). A Cells were lysed, and western blot analysis was conducted using specific antibodies as indicated (representative blot). B Quantification of band intensity of experiments as in A, calculated by the ImageJ software. The Y-axis represents the band intensity of β-catenin standardized to actin (β-catenin/actin) of Wnt-treated cells relative to control-treated cells (100%). C-E Treated monocytes (as in A) were stained and visualized using immunofluorescence. Images were obtained using a confocal microscope with a 60X oil-immersion objective lens C . An independent script was used to quantify the signal intensity and nuclear localization of β-catenin. Pairs signify individual donors, where each point represents the mean value of at least 5 fields D, E (see methods section). * P -value = 0.0359 for paired t-test of signal intensity D and * P -value = 0.0342 for paired t-test of nuclear localization E

    Article Snippet: L-Wnt-3a cells, L cells, and THP-1 cells were propagated according to the ATCC guidelines.

    Techniques: Expressing, Isolation, Cell Culture, Western Blot, Software, Staining, Immunofluorescence, Microscopy

    Wnt-3a does not induce expression of classical Wnt signaling target genes in monocytes. A Treated monocytes (as in Fig. ) were lysed for RT-qPCR analysis of the indicated Wnt target genes. B Cell protein lysates were analyzed by western blot using specific antibodies as indicated. The graphs represent the relative band intensity. C Treated monocytes were stained and visualized using immunofluorescence with the indicated antibodies. Signal intensity was quantified with no significant differences for any paired t-test

    Journal: Cell Communication and Signaling : CCS

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    doi: 10.1186/s12964-024-01608-8

    Figure Lengend Snippet: Wnt-3a does not induce expression of classical Wnt signaling target genes in monocytes. A Treated monocytes (as in Fig. ) were lysed for RT-qPCR analysis of the indicated Wnt target genes. B Cell protein lysates were analyzed by western blot using specific antibodies as indicated. The graphs represent the relative band intensity. C Treated monocytes were stained and visualized using immunofluorescence with the indicated antibodies. Signal intensity was quantified with no significant differences for any paired t-test

    Article Snippet: L-Wnt-3a cells, L cells, and THP-1 cells were propagated according to the ATCC guidelines.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining, Immunofluorescence

    Wnt-3a modifies the levels of monocyte-secreted immune proteins. Freshly isolated monocytes (from two donors) were cultured (in triplicates) for 8 h in the presence of control or Wnt-3a conditioned media (CM) for RNA sequencing. A Principal component analysis based on entire gene expression patterns, with batch effect removal to standardize the different donors. B Volcano plot of all genes based on differential expression analysis between control- and Wnt-treated monocytes. Significantly regulated genes are in red on either side (as defined by adjusted p -value < 0.05 and absolute fold change > 2). C, D Summary of Log2 Fold Change of significantly regulated cytokine and chemokine ligands C and receptors D . E Similarly treated monocytes from multiple blood samples were lysed for RT-qPCR assays of the indicated cytokines and chemokines

    Journal: Cell Communication and Signaling : CCS

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    doi: 10.1186/s12964-024-01608-8

    Figure Lengend Snippet: Wnt-3a modifies the levels of monocyte-secreted immune proteins. Freshly isolated monocytes (from two donors) were cultured (in triplicates) for 8 h in the presence of control or Wnt-3a conditioned media (CM) for RNA sequencing. A Principal component analysis based on entire gene expression patterns, with batch effect removal to standardize the different donors. B Volcano plot of all genes based on differential expression analysis between control- and Wnt-treated monocytes. Significantly regulated genes are in red on either side (as defined by adjusted p -value < 0.05 and absolute fold change > 2). C, D Summary of Log2 Fold Change of significantly regulated cytokine and chemokine ligands C and receptors D . E Similarly treated monocytes from multiple blood samples were lysed for RT-qPCR assays of the indicated cytokines and chemokines

    Article Snippet: L-Wnt-3a cells, L cells, and THP-1 cells were propagated according to the ATCC guidelines.

    Techniques: Isolation, Cell Culture, RNA Sequencing Assay, Expressing, Quantitative RT-PCR

    Wnt-3a affects chemokine secretion from monocytes. Media collected from the treated monocytes from two donors were centrifuged twice to remove the cells, and subjected to a chemokine membrane array (see methods section). Analysis of a representative membrane is shown in the two left panels. A summary of the changes in the chemokines detected in the two experiments is presented on the right: upregulated (red), downregulated (blue), and unchanged (grey) proteins. Original images and analyses are presented in Fig. S

    Journal: Cell Communication and Signaling : CCS

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    doi: 10.1186/s12964-024-01608-8

    Figure Lengend Snippet: Wnt-3a affects chemokine secretion from monocytes. Media collected from the treated monocytes from two donors were centrifuged twice to remove the cells, and subjected to a chemokine membrane array (see methods section). Analysis of a representative membrane is shown in the two left panels. A summary of the changes in the chemokines detected in the two experiments is presented on the right: upregulated (red), downregulated (blue), and unchanged (grey) proteins. Original images and analyses are presented in Fig. S

    Article Snippet: L-Wnt-3a cells, L cells, and THP-1 cells were propagated according to the ATCC guidelines.

    Techniques: Membrane

    Treating monocytes with Wnt-3a induces cell migration and CCL2 production. Naïve monocytes were plated in the top chamber of a transwell and exposed to different conditioned media (CM). Cells that crossed the transwell were collected from the bottom well and counted by flow cytometry. A Schematic illustration of experimental design. Naïve monocytes were exposed to Wnt-3a/control CM obtained from L-cells for 2 h (I). Naïve monocytes were exposed to Wnt-3a/control CM obtained from monocytes treated with Wnt-3a/control for 8 h (II). Treated monocytes (as in II) were washed and transferred to fresh RPMI medium for an additional 1 h. CM was collected and used to treat naïve monocytes in the transwell assay (III). B Flow cytometry counts of cells crossing the transwell. * P -value = 0.0307, *** P -value = 0.0003 and ** P -value = 0.0045 for paired t-tests. Y-axis represents the absolute cell count. C ELISA assays of CCL2 concentration in media used for the indicated experiments. Protein concentrations were calculated by regression from a standard curve. D Flow cytometry counts of cells crossing the transwell. Y-axis represents the absolute cell count. X-axis represents the media used as in I and II of the experimental design, with or without 2 μg/ml CCL2 inhibitor, marked as circles and squares, respectively. The white and black shapes represent each donor. FC – average fold change of the marked pairs. E A representative blot of media collected from the top and bottom chambers of the different experiments (as indicated), at the end of the transwell incubation period. Media were centrifuged to remove cells and analyzed by western blot using a specific anti-Wnt-3a antibody. C – control/control-treated media, W3 – Wnt-3a/Wnt-3a-treated media

    Journal: Cell Communication and Signaling : CCS

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    doi: 10.1186/s12964-024-01608-8

    Figure Lengend Snippet: Treating monocytes with Wnt-3a induces cell migration and CCL2 production. Naïve monocytes were plated in the top chamber of a transwell and exposed to different conditioned media (CM). Cells that crossed the transwell were collected from the bottom well and counted by flow cytometry. A Schematic illustration of experimental design. Naïve monocytes were exposed to Wnt-3a/control CM obtained from L-cells for 2 h (I). Naïve monocytes were exposed to Wnt-3a/control CM obtained from monocytes treated with Wnt-3a/control for 8 h (II). Treated monocytes (as in II) were washed and transferred to fresh RPMI medium for an additional 1 h. CM was collected and used to treat naïve monocytes in the transwell assay (III). B Flow cytometry counts of cells crossing the transwell. * P -value = 0.0307, *** P -value = 0.0003 and ** P -value = 0.0045 for paired t-tests. Y-axis represents the absolute cell count. C ELISA assays of CCL2 concentration in media used for the indicated experiments. Protein concentrations were calculated by regression from a standard curve. D Flow cytometry counts of cells crossing the transwell. Y-axis represents the absolute cell count. X-axis represents the media used as in I and II of the experimental design, with or without 2 μg/ml CCL2 inhibitor, marked as circles and squares, respectively. The white and black shapes represent each donor. FC – average fold change of the marked pairs. E A representative blot of media collected from the top and bottom chambers of the different experiments (as indicated), at the end of the transwell incubation period. Media were centrifuged to remove cells and analyzed by western blot using a specific anti-Wnt-3a antibody. C – control/control-treated media, W3 – Wnt-3a/Wnt-3a-treated media

    Article Snippet: L-Wnt-3a cells, L cells, and THP-1 cells were propagated according to the ATCC guidelines.

    Techniques: Migration, Flow Cytometry, Transwell Assay, Cell Counting, Enzyme-linked Immunosorbent Assay, Concentration Assay, Incubation, Western Blot

    The inflammatory state of patients with rheumatic joint diseases affects monocyte reaction to Wnt-3a stimulus. A Principal component analyses (PCA) based on RT-qPCR data (presented in Fig. S B) of Wnt-3a-treated monocytes (treated as in Fig. E). The blue X represents the centroid of the healthy donor samples, calculated as the average of the x and y coordinates of all the blue dots on the PCA graph. The bar graphs represent a calculation of distance of each dot from the healthy donor centroid, based on x,y coordinates. One-way ANOVA with Dunnett's multiple comparisons test to healthy control were employed. Healthy donors (blue), treated RJD patients (pink) and RJD patients naïve to treatment (maroon). ** P -value = 0.0024, ns = 0.6717. B Media were collected from the treated monocytes of two RJD patients naïve to immunosuppressive treatment. The media were subjected to a chemokine membrane array and analyzed, as performed with media of healthy donors in Fig. . The heatmap was created using numerical data of both the data of healthy donors represented in Fig. (Healthy 1&2) and the data of the two RJD patients (RJD 1&2). The fold change (FC) of signal ratio of Wnt-treated to control-treated monocyte is shown as a heatmap (log2(FC)), where upregulated genes are represented in red, downregulated in blue, and unchanged in white. Hierarchical clustering was performed on both the donors on the X-axis and the secreted proteins on the Y-axis. Only proteins clearly detected across all four membranes are represented. Original images and analyses of all 4 donors are presented in Fig. S , Fig. S , and the raw figure data (Supplementary file ). RJD – rheumatic joint diseases

    Journal: Cell Communication and Signaling : CCS

    Article Title: Wnt signaling regulates chemokine production and cell migration of circulating human monocytes

    doi: 10.1186/s12964-024-01608-8

    Figure Lengend Snippet: The inflammatory state of patients with rheumatic joint diseases affects monocyte reaction to Wnt-3a stimulus. A Principal component analyses (PCA) based on RT-qPCR data (presented in Fig. S B) of Wnt-3a-treated monocytes (treated as in Fig. E). The blue X represents the centroid of the healthy donor samples, calculated as the average of the x and y coordinates of all the blue dots on the PCA graph. The bar graphs represent a calculation of distance of each dot from the healthy donor centroid, based on x,y coordinates. One-way ANOVA with Dunnett's multiple comparisons test to healthy control were employed. Healthy donors (blue), treated RJD patients (pink) and RJD patients naïve to treatment (maroon). ** P -value = 0.0024, ns = 0.6717. B Media were collected from the treated monocytes of two RJD patients naïve to immunosuppressive treatment. The media were subjected to a chemokine membrane array and analyzed, as performed with media of healthy donors in Fig. . The heatmap was created using numerical data of both the data of healthy donors represented in Fig. (Healthy 1&2) and the data of the two RJD patients (RJD 1&2). The fold change (FC) of signal ratio of Wnt-treated to control-treated monocyte is shown as a heatmap (log2(FC)), where upregulated genes are represented in red, downregulated in blue, and unchanged in white. Hierarchical clustering was performed on both the donors on the X-axis and the secreted proteins on the Y-axis. Only proteins clearly detected across all four membranes are represented. Original images and analyses of all 4 donors are presented in Fig. S , Fig. S , and the raw figure data (Supplementary file ). RJD – rheumatic joint diseases

    Article Snippet: L-Wnt-3a cells, L cells, and THP-1 cells were propagated according to the ATCC guidelines.

    Techniques: Quantitative RT-PCR, Membrane