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sheep fap  (R&D Systems)


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    R&D Systems sheep fap
    Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein <t>(FAP)</t> and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained <t>unchanged.</t> <t>TLR4</t> expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).
    Sheep Fap, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 95 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sheep+anti+human+fap/Human+Fibroblast+Activation+Protein+alpha%2FFAP+Antibody/pmc12733209-236-52-55
    Average 95 stars, based on 95 article reviews
    sheep fap - by Bioz Stars, 2026-09
    95/100 stars

    Images

    1) Product Images from "BGN Secreted by Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression via Activation of TLR4-Mediated Erk and NF-κB Signaling Pathways"

    Article Title: BGN Secreted by Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression via Activation of TLR4-Mediated Erk and NF-κB Signaling Pathways

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms262412024

    Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein (FAP) and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained unchanged. TLR4 expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).
    Figure Legend Snippet: Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein (FAP) and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained unchanged. TLR4 expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).

    Techniques Used: Migration, Activation Assay, Expressing, RNA Sequencing, MTS Assay, Transwell Migration Assay, Recombinant, Western Blot, Phospho-proteomics

    Related Articles

    Immunostaining:

    Article Title: CCL18 signaling from tumor-associated macrophages activates fibroblasts to adopt a chemoresistance-inducing phenotype
    Article Snippet: .. For immunostaining of cultured cells, the cells were fixed with paraformaldehyde and then permeabilized with 0.1% Triton X-100 on ice for 15 min. Nonspecific antigen epitope binding was blocked by incubation with phosphate buffer containing 5% BSA for 1 h. Next, sections or cells were probed overnight at 4 °C with specific primary antibodies, including goat anti-human α-SMA (Abcam, # ab21027, 1:100), rabbit anti-human α-SMA (Abcam, # ab124964, 1:100), mouse anti-human ALDH1 (R&D, # AF5869, 1:100), rabbit anti-human CD10 (Abcam, # ab73409, 1:30), mouse anti-human GPR77 (BioLegend, #342402, 1:30), sheep anti-human FAP (R&D, # AF3715, 1:100), rabbit anti-human CD68 (Abcam, # ab213363, 1:100), goat anti-human CCL18 (R&D, # AF394, 1:100), rabbit anti-human p65 (CST, # 8242, 1:50), rabbit anti-human ac-p65 (Abcam, # ab19870, 1:50), goat anti-human IL6 (R&D, # AF-206, 10 μg/mL) and mouse anti-human IL8 (R&D, # MAB208, 20 μg/mL). .. Then, antigen–antibody binding was visualized by using Alexa Fluor-conjugated secondary antibodies (Invitrogen) according to the manufacturer’s instructions.

    Cell Culture:

    Article Title: CCL18 signaling from tumor-associated macrophages activates fibroblasts to adopt a chemoresistance-inducing phenotype
    Article Snippet: .. For immunostaining of cultured cells, the cells were fixed with paraformaldehyde and then permeabilized with 0.1% Triton X-100 on ice for 15 min. Nonspecific antigen epitope binding was blocked by incubation with phosphate buffer containing 5% BSA for 1 h. Next, sections or cells were probed overnight at 4 °C with specific primary antibodies, including goat anti-human α-SMA (Abcam, # ab21027, 1:100), rabbit anti-human α-SMA (Abcam, # ab124964, 1:100), mouse anti-human ALDH1 (R&D, # AF5869, 1:100), rabbit anti-human CD10 (Abcam, # ab73409, 1:30), mouse anti-human GPR77 (BioLegend, #342402, 1:30), sheep anti-human FAP (R&D, # AF3715, 1:100), rabbit anti-human CD68 (Abcam, # ab213363, 1:100), goat anti-human CCL18 (R&D, # AF394, 1:100), rabbit anti-human p65 (CST, # 8242, 1:50), rabbit anti-human ac-p65 (Abcam, # ab19870, 1:50), goat anti-human IL6 (R&D, # AF-206, 10 μg/mL) and mouse anti-human IL8 (R&D, # MAB208, 20 μg/mL). .. Then, antigen–antibody binding was visualized by using Alexa Fluor-conjugated secondary antibodies (Invitrogen) according to the manufacturer’s instructions.

    Binding Assay:

    Article Title: CCL18 signaling from tumor-associated macrophages activates fibroblasts to adopt a chemoresistance-inducing phenotype
    Article Snippet: .. For immunostaining of cultured cells, the cells were fixed with paraformaldehyde and then permeabilized with 0.1% Triton X-100 on ice for 15 min. Nonspecific antigen epitope binding was blocked by incubation with phosphate buffer containing 5% BSA for 1 h. Next, sections or cells were probed overnight at 4 °C with specific primary antibodies, including goat anti-human α-SMA (Abcam, # ab21027, 1:100), rabbit anti-human α-SMA (Abcam, # ab124964, 1:100), mouse anti-human ALDH1 (R&D, # AF5869, 1:100), rabbit anti-human CD10 (Abcam, # ab73409, 1:30), mouse anti-human GPR77 (BioLegend, #342402, 1:30), sheep anti-human FAP (R&D, # AF3715, 1:100), rabbit anti-human CD68 (Abcam, # ab213363, 1:100), goat anti-human CCL18 (R&D, # AF394, 1:100), rabbit anti-human p65 (CST, # 8242, 1:50), rabbit anti-human ac-p65 (Abcam, # ab19870, 1:50), goat anti-human IL6 (R&D, # AF-206, 10 μg/mL) and mouse anti-human IL8 (R&D, # MAB208, 20 μg/mL). .. Then, antigen–antibody binding was visualized by using Alexa Fluor-conjugated secondary antibodies (Invitrogen) according to the manufacturer’s instructions.

    Incubation:

    Article Title: CCL18 signaling from tumor-associated macrophages activates fibroblasts to adopt a chemoresistance-inducing phenotype
    Article Snippet: .. For immunostaining of cultured cells, the cells were fixed with paraformaldehyde and then permeabilized with 0.1% Triton X-100 on ice for 15 min. Nonspecific antigen epitope binding was blocked by incubation with phosphate buffer containing 5% BSA for 1 h. Next, sections or cells were probed overnight at 4 °C with specific primary antibodies, including goat anti-human α-SMA (Abcam, # ab21027, 1:100), rabbit anti-human α-SMA (Abcam, # ab124964, 1:100), mouse anti-human ALDH1 (R&D, # AF5869, 1:100), rabbit anti-human CD10 (Abcam, # ab73409, 1:30), mouse anti-human GPR77 (BioLegend, #342402, 1:30), sheep anti-human FAP (R&D, # AF3715, 1:100), rabbit anti-human CD68 (Abcam, # ab213363, 1:100), goat anti-human CCL18 (R&D, # AF394, 1:100), rabbit anti-human p65 (CST, # 8242, 1:50), rabbit anti-human ac-p65 (Abcam, # ab19870, 1:50), goat anti-human IL6 (R&D, # AF-206, 10 μg/mL) and mouse anti-human IL8 (R&D, # MAB208, 20 μg/mL). .. Then, antigen–antibody binding was visualized by using Alexa Fluor-conjugated secondary antibodies (Invitrogen) according to the manufacturer’s instructions.

    Immunohistochemistry:

    Article Title: Human Pancreatic Tumor Organoids Reveal Loss of Stem Cell Niche Factor Dependence during Disease Progression.
    Article Snippet: .. For immunohistochemistry, rat anti-integrin-a6 (313602, Biolegend, 1:100), mouse anti-aSMA (#MS-113-P, Thermo Fisher Scientific, 1:800), sheep anti-human FAP (#AF3715, R&D systems, 1:50), rabbit anti-GATA6 (#5851, CST, 1:300), anti-cytokeratin19 (#NCL-L-CK19, Novocastra, 1:100) were used. .. The secondary antibodies were horseradish peroxidase-conjugated anti-rabbit antibodies (Nichirei Bioscience) or Alexa Fluor 568-conjugated anti-rat, mouse, sheep antibodies (Thermo Scientific or Abcam).

    Flow Cytometry:

    Article Title: FAP-retargeted Ad5 enables in vivo gene delivery to stromal cells in the tumor microenvironment
    Article Snippet: Flow cytometry analysis was performed using FlowJoTM software (BD Biosciences). .. Antibodies used for flow cytometry included: mouse anti-human FAP-APC (R&D Systems, FAB3715A), sheep anti-human FAP (R&D Systems, AF3715), mouse anti-mouse FAP (Merck, MABC1145), mouse anti-CAR (Millipore, 05-644), mouse anti-HER2-AF488 (BioLegend, 324410), mouse anti-HER2-AF647 (BioLegend, 324412), rat anti-mouse CD45-AF700 (BioLegend, 103127), donkey anti-sheep IgG-CF488A (Sigma-Aldrich, SAB4600038), goat anti-mouse IgG-AF647 (Invitrogen, A21235), goat anti-mouse IgG-AF488 (Invitrogen, A11001). ..



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    Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein <t>(FAP)</t> and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained <t>unchanged.</t> <t>TLR4</t> expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).
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    93
    R&D Systems biotinylated polyclonal sheep anti human fap antibody
    Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein <t>(FAP)</t> and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained <t>unchanged.</t> <t>TLR4</t> expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).
    Biotinylated Polyclonal Sheep Anti Human Fap Antibody, 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
    https://www.bioz.com/product/sheep+anti+human+fap/Human+Fibroblast+Activation+Protein+alpha%2FFAP+Biotinylated+Antibody/pmc11346374-113-3-11
    Average 93 stars, based on 1 article reviews
    biotinylated polyclonal sheep anti human fap antibody - by Bioz Stars, 2026-09
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    sheep anti human fap antibody  (R&D Systems)
    95
    R&D Systems sheep anti human fap antibody
    Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein <t>(FAP)</t> and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained <t>unchanged.</t> <t>TLR4</t> expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).
    Sheep Anti Human Fap Antibody, 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
    https://www.bioz.com/product/sheep+anti+human+fap/Human+Fibroblast+Activation+Protein+alpha%2FFAP+Antibody/pmc11324301__jci___134___173835___s067-17-15-21
    Average 95 stars, based on 1 article reviews
    sheep anti human fap antibody - by Bioz Stars, 2026-09
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    sheep antibody against fap  (R&D Systems)
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    Figure 1 Esophageal squamous cell carcinoma (ESCC) cells exhibit enhanced survival, growth, and migration after direct co-culture with mesenchymal stem cells (MSCs). A: Direct co-culture of ESCC cell lines (TE-9, TE-10, and TE-15) and MSCs was performed for 4 days. MSCs and TE cells were seeded similarly without co-culture and cultured for 4 days as mono-cultured controls. After direct co-culture, epithelial cell adhesion <t>molecule</t> <t>(EpCAM)epositive</t> (TE-9 co, TE-10 co, and TE-15 co) and EpCAM-negative cells [cancer-associated fibroblast (CAF) 9, CAF10, and CAF15] were separated through magnetic-activated cell sorting (MACS). Mono-cultured MSCs (MSC mono) and mono-cultured TE cells (TE-9 mono, TE-10 mono, and TE-15 mono) were passed through MACS as controls. B: In each cell after separation using MACS, RT-PCR confirmed the expression of EPCAM, cadherin 1 (CDH1), fibroblast activation protein <t>(FAP),</t> and actin alpha 2 (ACTA2) (left panel), and Western blot analysis confirmed the expression of EpCAM, E-cadherin, FAP, and a-smooth muscle actin (a-SMA) (right panel). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta actin (ACTB) were used as controls in RT-PCR and Western blot analysis, respectively. C: FAP expression in CAF9, CAF10, and CAF15 was compared with that in MSCs using real-time quantitative PCR. D: MTS assays were performed between TE cells after mono-culture and direct co-culture to evaluate the survival and growth of TE cells. E: Transwell migration assays were performed between TE cells after mono-culture and direct co-culture. Migrating cells were counted in five random fields in each chamber after 48 hours of incubation. Typical images are shown below. F: Western blot analysis of Akt, phosphorylated Akt (p-Akt; Ser473), p-Akt (Thr308), extracellular signal-regulated kinase (Erk), and phosphorylated Erk (p-Erk; Thr202/Tyr204) in mono-cultured and co-cultured TE-9, TE-10, TE-15 cells and MSC mono, CAF9, CAF10, and CAF15. ACTB was used as a control. Data are presented as means SEM (CeE). *P < 0.05, ***P < 0.001. Scale bars Z 100 mm (E).
    Sheep Antibody Against Fap, 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
    https://www.bioz.com/product/sheep+anti+human+fap/Human+Fibroblast+Activation+Protein+alpha%2FFAP+Antibody/pm38320632-55-20-27
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    Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein (FAP) and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained unchanged. TLR4 expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).

    Journal: International Journal of Molecular Sciences

    Article Title: BGN Secreted by Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression via Activation of TLR4-Mediated Erk and NF-κB Signaling Pathways

    doi: 10.3390/ijms262412024

    Figure Lengend Snippet: Biglycan (BGN) promotes proliferation, migration, and activation of macrophages and fibroblasts. ( A ) Cell–cell communication network analysis of BGN -expressing cancer-associated fibroblasts (CAFs) in esophageal squamous cell carcinoma (ESCC) tissues. Fibroblasts in single-cell RNA sequencing (scRNAseq) datasets from ESCC tissues were stratified into CAF_ BGN _High (scaled expression > 3) and CAF_ BGN _Low (≤3) groups based on BGN expression levels. Cell–cell communication inferred by CellChat revealed that CAF_ BGN _High cells exhibited strong outgoing signaling toward epithelial and myeloid cells, as well as prominent autocrine signaling within the CAF_ BGN _High population. ( B , C ) MTS assay ( B ) and Transwell migration assay ( C ) showing increased proliferation and migration of mesenchymal stem cells (MSCs) treated with recombinant human BGN (rhBGN; 100 ng/mL), respectively. ( D ) Western Blot analysis showing that fibroblast activation protein (FAP) and α-smooth muscle actin (αSMA) (CAF markers) were upregulated in MSCs treated with rhBGN for 24 h, whereas interleukin-6 (IL6) expression remained unchanged. TLR4 expression was detected; however, phosphorylation of NF-κB and Erk was not increased by rhBGN. ( E , F ) MTS assay ( E ) and Transwell migration assay ( F ) showing that rhBGN-induced proliferation and migration of MSCs were not suppressed by a TLR4-neutralizing antibody (1 μg/mL). ( G , H ) MTS assay ( G ) and Transwell migration assay ( H ) showing increased proliferation and migration of macrophages treated with rhBGN (100 ng/mL), respectively. ( I ) Western Blot analysis showing that rhBGN treatment upregulated the expression of CD163 and CD206 (M2 macrophage markers) and increased NF-κB phosphorylation, while TLR4 expression was also detected, but phosphorylated Erk was not changed. ( J , K ) MTS assay ( J ) and Transwell migration assay ( K ) showing that rhBGN-induced proliferation and migration of macrophages were attenuated by treatment with a TLR4-neutralizing antibody (1 μg/mL), respectively. ( L , M ) MTS assay ( L ) and Transwell migration assay ( M ) showing that treatment with the NF-κB pathway inhibitor Bay 11-7082 (1 μM) attenuated rhBGN-induced proliferation and migration to a similar extent, respectively. Data are presented as the mean ± standard error of the mean (SEM) from three independent experiments ( B , C , E – H , J – M ). * p < 0.05, ** p < 0.01, *** p < 0.001. N.S., not significant. Scale bars: 100 μm ( C , F , H , K , M ).

    Article Snippet: The primary antibodies used as follows: rabbit BGN (#16409-1-AP, R&D Systems), rabbit phosphorylated Erk1/2 (pErk; #9101, Cell signaling Technology; CST, Danvers, MA, USA), rabbit total Erk1/2 (tErk; #9102, CST), rabbit phosphorylated NF-κB p65 (pNF-κB; #3033, CST), rabbit total NF-κB p65 (tNF-κB; #8242, CST), mouse TLR4 (#sc-293072, Santa Cruz Biotechmology, Dallas, TX, USA), sheep FAP (#AF3715, R&D Systems), rabbit αSMA (#ab5694, Abcam, Cambridge, UK), rabbit IL-6 (#ab6672, Abcam), mouse CD163 (#NCL-L-CD163; Leica Biosystems, Wetzlar, Germany), mouse CD206 (#sc-376108, Santa Cruz), and rabbit β-actin (#4970, CST) antibodies.

    Techniques: Migration, Activation Assay, Expressing, RNA Sequencing, MTS Assay, Transwell Migration Assay, Recombinant, Western Blot, Phospho-proteomics

    Figure 1 Esophageal squamous cell carcinoma (ESCC) cells exhibit enhanced survival, growth, and migration after direct co-culture with mesenchymal stem cells (MSCs). A: Direct co-culture of ESCC cell lines (TE-9, TE-10, and TE-15) and MSCs was performed for 4 days. MSCs and TE cells were seeded similarly without co-culture and cultured for 4 days as mono-cultured controls. After direct co-culture, epithelial cell adhesion molecule (EpCAM)epositive (TE-9 co, TE-10 co, and TE-15 co) and EpCAM-negative cells [cancer-associated fibroblast (CAF) 9, CAF10, and CAF15] were separated through magnetic-activated cell sorting (MACS). Mono-cultured MSCs (MSC mono) and mono-cultured TE cells (TE-9 mono, TE-10 mono, and TE-15 mono) were passed through MACS as controls. B: In each cell after separation using MACS, RT-PCR confirmed the expression of EPCAM, cadherin 1 (CDH1), fibroblast activation protein (FAP), and actin alpha 2 (ACTA2) (left panel), and Western blot analysis confirmed the expression of EpCAM, E-cadherin, FAP, and a-smooth muscle actin (a-SMA) (right panel). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta actin (ACTB) were used as controls in RT-PCR and Western blot analysis, respectively. C: FAP expression in CAF9, CAF10, and CAF15 was compared with that in MSCs using real-time quantitative PCR. D: MTS assays were performed between TE cells after mono-culture and direct co-culture to evaluate the survival and growth of TE cells. E: Transwell migration assays were performed between TE cells after mono-culture and direct co-culture. Migrating cells were counted in five random fields in each chamber after 48 hours of incubation. Typical images are shown below. F: Western blot analysis of Akt, phosphorylated Akt (p-Akt; Ser473), p-Akt (Thr308), extracellular signal-regulated kinase (Erk), and phosphorylated Erk (p-Erk; Thr202/Tyr204) in mono-cultured and co-cultured TE-9, TE-10, TE-15 cells and MSC mono, CAF9, CAF10, and CAF15. ACTB was used as a control. Data are presented as means SEM (CeE). *P < 0.05, ***P < 0.001. Scale bars Z 100 mm (E).

    Journal: The American journal of pathology

    Article Title: Periostin in Cancer-Associated Fibroblasts Promotes Esophageal Squamous Cell Carcinoma Progression by Enhancing Cancer and Stromal Cell Migration.

    doi: 10.1016/j.ajpath.2023.12.010

    Figure Lengend Snippet: Figure 1 Esophageal squamous cell carcinoma (ESCC) cells exhibit enhanced survival, growth, and migration after direct co-culture with mesenchymal stem cells (MSCs). A: Direct co-culture of ESCC cell lines (TE-9, TE-10, and TE-15) and MSCs was performed for 4 days. MSCs and TE cells were seeded similarly without co-culture and cultured for 4 days as mono-cultured controls. After direct co-culture, epithelial cell adhesion molecule (EpCAM)epositive (TE-9 co, TE-10 co, and TE-15 co) and EpCAM-negative cells [cancer-associated fibroblast (CAF) 9, CAF10, and CAF15] were separated through magnetic-activated cell sorting (MACS). Mono-cultured MSCs (MSC mono) and mono-cultured TE cells (TE-9 mono, TE-10 mono, and TE-15 mono) were passed through MACS as controls. B: In each cell after separation using MACS, RT-PCR confirmed the expression of EPCAM, cadherin 1 (CDH1), fibroblast activation protein (FAP), and actin alpha 2 (ACTA2) (left panel), and Western blot analysis confirmed the expression of EpCAM, E-cadherin, FAP, and a-smooth muscle actin (a-SMA) (right panel). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta actin (ACTB) were used as controls in RT-PCR and Western blot analysis, respectively. C: FAP expression in CAF9, CAF10, and CAF15 was compared with that in MSCs using real-time quantitative PCR. D: MTS assays were performed between TE cells after mono-culture and direct co-culture to evaluate the survival and growth of TE cells. E: Transwell migration assays were performed between TE cells after mono-culture and direct co-culture. Migrating cells were counted in five random fields in each chamber after 48 hours of incubation. Typical images are shown below. F: Western blot analysis of Akt, phosphorylated Akt (p-Akt; Ser473), p-Akt (Thr308), extracellular signal-regulated kinase (Erk), and phosphorylated Erk (p-Erk; Thr202/Tyr204) in mono-cultured and co-cultured TE-9, TE-10, TE-15 cells and MSC mono, CAF9, CAF10, and CAF15. ACTB was used as a control. Data are presented as means SEM (CeE). *P < 0.05, ***P < 0.001. Scale bars Z 100 mm (E).

    Article Snippet: The primary antibodies were as follows: mouse antibody against EpCAM (1:100; number 2929); rabbit antibody against Ecadherin (1:100; number 3159); sheep antibody against FAP (1:300; number AF3715; R&D Systems); rabbit antibody against a-smooth muscle actin (1:300; number ab5694; Sigma-Aldrich); rabbit antibody against phosphorylated Akt (p-Akt; Ser473; 1:100; number 4060); rabbit antibody against phosphorylated Akt (Thr308; 1:100; number 2965); rabbit antibody against Akt (1:100; number 9272); rabbit antibody against phosphorylated extracellular signalregulated kinase (Erk; Thr202/Tyr204; 1:100; number 9101); rabbit antibody against Erk (1:100; number 9102); rabbit antibody against integrin b4 (1:200; number 14803); mouse antibody against CD163 (1:100; number NCL-LCD163; Leica Biosystems, Wetzlar, Germany); mouse antibody against CD204 (1:200; number KT022; Trans Genic, Kobe, Japan); and rabbit antibody against ACTB (1:2000; number 4970).

    Techniques: Migration, Co-Culture Assay, Cell Culture, FACS, Reverse Transcription Polymerase Chain Reaction, Expressing, Activation Assay, Western Blot, Real-time Polymerase Chain Reaction, Incubation, Control