tgf β1  (R&D Systems)

 
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  • 97
    Name:
    Human TGF beta 1 Protein CF
    Description:
    The Human TGF beta 1 Protein from R D Systems is derived from Human Platelets The Human TGF beta 1 Protein has been validated for the following applications Bioactivity
    Catalog Number:
    100-B-001/CF
    Price:
    289
    Category:
    Proteins and Enzymes
    Source:
    Human Platelets
    Applications:
    Bioactivity
    Purity:
    >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
    Conjugate:
    Unconjugated
    Size:
    1 ug
    Buy from Supplier


    Structured Review

    R&D Systems tgf β1
    Human TGF beta 1 Protein CF
    The Human TGF beta 1 Protein from R D Systems is derived from Human Platelets The Human TGF beta 1 Protein has been validated for the following applications Bioactivity
    https://www.bioz.com/result/tgf β1/product/R&D Systems
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tgf β1 - by Bioz Stars, 2021-05
    97/100 stars

    Images

    1) Product Images from "Differential Expression of TGF-β Isoforms in Human Kerationocytes by Narrow Band UVB"

    Article Title: Differential Expression of TGF-β Isoforms in Human Kerationocytes by Narrow Band UVB

    Journal: Annals of Dermatology

    doi: 10.5021/ad.2008.20.3.113

    TGF-β1 protein levels at 800, 1,000 and 1,200 mJ/cm 2 for 24 hr and 48 hr when compared with the control group. TGF-β1 protein levels in supernatant was determined by ELISA assay (duplicates). Values are the mean±SD of 6 samples
    Figure Legend Snippet: TGF-β1 protein levels at 800, 1,000 and 1,200 mJ/cm 2 for 24 hr and 48 hr when compared with the control group. TGF-β1 protein levels in supernatant was determined by ELISA assay (duplicates). Values are the mean±SD of 6 samples

    Techniques Used: Enzyme-linked Immunosorbent Assay

    TGF-β1 mRNA levels at 800, 1,000 and 1,200 mJ/cm 2 compared with control group for 24 hr and 48 hr. (A) Representative RT-PCR showing TGF-β1 mRNA in all samples. (B) Graphical representation of combined densitometric analyses of TGF-β1
    Figure Legend Snippet: TGF-β1 mRNA levels at 800, 1,000 and 1,200 mJ/cm 2 compared with control group for 24 hr and 48 hr. (A) Representative RT-PCR showing TGF-β1 mRNA in all samples. (B) Graphical representation of combined densitometric analyses of TGF-β1

    Techniques Used: Reverse Transcription Polymerase Chain Reaction

    2) Product Images from "The A2B Adenosine Receptor Promotes Th17 Differentiation via Stimulation of Dendritic Cell IL-6"

    Article Title: The A2B Adenosine Receptor Promotes Th17 Differentiation via Stimulation of Dendritic Cell IL-6

    Journal: Journal of Immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1100117

    An adenosine mimetic favors Th17 cells in the presence of TGF-β1. Naive CD4 + T cells were activated with BMDCs and anti-CD3 in the presence of TGF-β1 and 500 nM NECA or vehicle control. Day 4 cells were harvested, washed, and replated in complete media overnight prior to restimulation with PMA/ionomycin. Dot plots show intracellular IL-17 and Foxp3 in CD4 + T cells. Dot plots ( A ) are representative of three or more independent experiments, and compiled data ( B ) were pooled from three or more independent experiments and represent the mean ± SEM. * p
    Figure Legend Snippet: An adenosine mimetic favors Th17 cells in the presence of TGF-β1. Naive CD4 + T cells were activated with BMDCs and anti-CD3 in the presence of TGF-β1 and 500 nM NECA or vehicle control. Day 4 cells were harvested, washed, and replated in complete media overnight prior to restimulation with PMA/ionomycin. Dot plots show intracellular IL-17 and Foxp3 in CD4 + T cells. Dot plots ( A ) are representative of three or more independent experiments, and compiled data ( B ) were pooled from three or more independent experiments and represent the mean ± SEM. * p

    Techniques Used:

    3) Product Images from "MiR-155-mediated loss of C/EBP? shifts the TGF-? response from growth inhibition to epithelial-mesenchymal transition, invasion and metastasis in breast cancer"

    Article Title: MiR-155-mediated loss of C/EBP? shifts the TGF-? response from growth inhibition to epithelial-mesenchymal transition, invasion and metastasis in breast cancer

    Journal: Oncogene

    doi: 10.1038/onc.2013.322

    Loss of C/EBPβ shifts the TGF-β response from growth inhibition to EMT. ( a ) Immunoblotting results showing the effect of knockdown of C/EBPβ via overexpression of Cebpb shRNA ( Cebpb shRNA) on the expression of EMT markers at baseline and after treatment with TGF-β1 (10 ng/ml, 48 h). Non-targeting shRNA was used as a control (Control shRNA). ( b ) qPCR results showing the effect of transient overexpression of LAP2 on Cdh1 and Cxadr expression in NMuMG cells at baseline, and after TGF-β1 treatment (2 ng/ml, 24 h). Data from qPCR experiments ( f , g ) represent mean±s.e.m. of three independent experiments. * P
    Figure Legend Snippet: Loss of C/EBPβ shifts the TGF-β response from growth inhibition to EMT. ( a ) Immunoblotting results showing the effect of knockdown of C/EBPβ via overexpression of Cebpb shRNA ( Cebpb shRNA) on the expression of EMT markers at baseline and after treatment with TGF-β1 (10 ng/ml, 48 h). Non-targeting shRNA was used as a control (Control shRNA). ( b ) qPCR results showing the effect of transient overexpression of LAP2 on Cdh1 and Cxadr expression in NMuMG cells at baseline, and after TGF-β1 treatment (2 ng/ml, 24 h). Data from qPCR experiments ( f , g ) represent mean±s.e.m. of three independent experiments. * P

    Techniques Used: Inhibition, Over Expression, shRNA, Expressing, Real-time Polymerase Chain Reaction

    Loss of C/EBPβ promotes invasion and metastatic spread of mammary tumor cells. ( a ) Immunoblotting results showing the effect of overexpression of Cebpb shRNA on the expression of E-cadherin, vimentin and C/EBPβ in 4T1 cells. Calnexin was used as a loading control. ( b ) Bar graph showing the effect of C/EBPβ knockdown through expression of Cebpb shRNA on the capacity of 4T1 cells to migrate through Matrigel using TGF-β1 (10 ng/ml) as a chemoattractant. Data are presented as percentage values compared with controls that were set to 100. ** P
    Figure Legend Snippet: Loss of C/EBPβ promotes invasion and metastatic spread of mammary tumor cells. ( a ) Immunoblotting results showing the effect of overexpression of Cebpb shRNA on the expression of E-cadherin, vimentin and C/EBPβ in 4T1 cells. Calnexin was used as a loading control. ( b ) Bar graph showing the effect of C/EBPβ knockdown through expression of Cebpb shRNA on the capacity of 4T1 cells to migrate through Matrigel using TGF-β1 (10 ng/ml) as a chemoattractant. Data are presented as percentage values compared with controls that were set to 100. ** P

    Techniques Used: Over Expression, shRNA, Expressing

    Loss of C/EBPβ is linked to the induction of EMT in response to TGF-β1. ( a , c ) Representative brightfield and immunofluorescence images showing induction of morphological characteristics of EMT and loss of nuclear staining of C/EBPβ in NMuMG cells ( a ) but not in EpH4 cells ( c ) after TGF-β1 treatment (10 ng/ml, 48 h). Scale bars, 50 μm. ( b , d ) Results from immunoblotting analysis showing changes in the expression of established EMT markers, and decreased expression of all three isoforms of C/EBPβ (LAP1, LAP2 and LIP) in NMuMG cells ( b ) but not in EpH4 cells ( d ) after TGF-β1 treatment (10 ng/ml, 48 h). ( e ) Immunoblotting analysis of the effect of TGF-β1 treatment (10 ng/ml, 1 h) on phosphorylation of Smad3 (p-Smad3) in NMuMG and EpH4 cells. ( f ) Immunoblotting analysis of the effect of siRNA-mediated knockdown of Smad3 (100 n M , 72 h) on the repression of C/EBPβ during TGF-β1-induced EMT (5 ng/ml, 24 h) in NMuMG cells. Calnexin was used as a loading control for all immunoblotting experiments.
    Figure Legend Snippet: Loss of C/EBPβ is linked to the induction of EMT in response to TGF-β1. ( a , c ) Representative brightfield and immunofluorescence images showing induction of morphological characteristics of EMT and loss of nuclear staining of C/EBPβ in NMuMG cells ( a ) but not in EpH4 cells ( c ) after TGF-β1 treatment (10 ng/ml, 48 h). Scale bars, 50 μm. ( b , d ) Results from immunoblotting analysis showing changes in the expression of established EMT markers, and decreased expression of all three isoforms of C/EBPβ (LAP1, LAP2 and LIP) in NMuMG cells ( b ) but not in EpH4 cells ( d ) after TGF-β1 treatment (10 ng/ml, 48 h). ( e ) Immunoblotting analysis of the effect of TGF-β1 treatment (10 ng/ml, 1 h) on phosphorylation of Smad3 (p-Smad3) in NMuMG and EpH4 cells. ( f ) Immunoblotting analysis of the effect of siRNA-mediated knockdown of Smad3 (100 n M , 72 h) on the repression of C/EBPβ during TGF-β1-induced EMT (5 ng/ml, 24 h) in NMuMG cells. Calnexin was used as a loading control for all immunoblotting experiments.

    Techniques Used: Immunofluorescence, Staining, Expressing

    Depletion of C/EBPβ during TGF-β1-induced EMT by miR-155. ( a ) Results from qPCR analysis of the effect of TGF-β1 (10 ng/ml, 24 h) on mRNA levels of Cebpb , Cdh1 , Cxadr and Vim relative to control, which was normalized to 100%. Data are mean ±s.e.m. of three independent experiments. ( b , c ) Immunoblotting analysis of the effect of a miR-155 inhibitor ( b ) and a miR-155 mimic ( c ) on the repression of C/EBPβ during TGF-β1-induced EMT (5 ng/ml, 24 h). ( d ) Results from qPCR analysis of the expression of miR-155 in EpH4 and NMuMG cells at baseline and after treatment with TGF-β1 (10 ng/ml, 24 h). ( e ) Immunoblotting analysis of the relative expression levels of C/EBPβ in EpH4 and NMuMG cells at baseline and after treatment with TGF-β1 (10 ng/ml, 48 h). Calnexin was used as a loading control for all immunoblotting experiments. * P
    Figure Legend Snippet: Depletion of C/EBPβ during TGF-β1-induced EMT by miR-155. ( a ) Results from qPCR analysis of the effect of TGF-β1 (10 ng/ml, 24 h) on mRNA levels of Cebpb , Cdh1 , Cxadr and Vim relative to control, which was normalized to 100%. Data are mean ±s.e.m. of three independent experiments. ( b , c ) Immunoblotting analysis of the effect of a miR-155 inhibitor ( b ) and a miR-155 mimic ( c ) on the repression of C/EBPβ during TGF-β1-induced EMT (5 ng/ml, 24 h). ( d ) Results from qPCR analysis of the expression of miR-155 in EpH4 and NMuMG cells at baseline and after treatment with TGF-β1 (10 ng/ml, 24 h). ( e ) Immunoblotting analysis of the relative expression levels of C/EBPβ in EpH4 and NMuMG cells at baseline and after treatment with TGF-β1 (10 ng/ml, 48 h). Calnexin was used as a loading control for all immunoblotting experiments. * P

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    4) Product Images from "Altered fibroblast proteoglycan production in COPD"

    Article Title: Altered fibroblast proteoglycan production in COPD

    Journal: Respiratory Research

    doi: 10.1186/1465-9921-11-55

    Proteoglycan production after TGF-β1 stimulation . Fibroblasts were incubated for 24 hours in cell medium containing 0.4% serum and 10 ng/ml TGF-β 1 , and proteoglycan content in medium was determined by the ability to incorporate [S 35 ]-sulphate into glycosoaminoglycan side chains. Values were related to protein concentration in the respective cell layers to account for variability in cell numbers. Individual proteoglycans were separated and quantified on SDS-PAGE gels to determine the contribution of each to total proteoglycan content. Each point represents the fold change after TGF-β 1 stimulation compared to basal levels. Dashed lines show the basal level of production of each proteoglycan. Graph A shows relative change of versican, B perlecan, C biglycan, and D decorin. * P
    Figure Legend Snippet: Proteoglycan production after TGF-β1 stimulation . Fibroblasts were incubated for 24 hours in cell medium containing 0.4% serum and 10 ng/ml TGF-β 1 , and proteoglycan content in medium was determined by the ability to incorporate [S 35 ]-sulphate into glycosoaminoglycan side chains. Values were related to protein concentration in the respective cell layers to account for variability in cell numbers. Individual proteoglycans were separated and quantified on SDS-PAGE gels to determine the contribution of each to total proteoglycan content. Each point represents the fold change after TGF-β 1 stimulation compared to basal levels. Dashed lines show the basal level of production of each proteoglycan. Graph A shows relative change of versican, B perlecan, C biglycan, and D decorin. * P

    Techniques Used: Incubation, Protein Concentration, SDS Page

    5) Product Images from "Two Distinct Regions of Latency-associated Peptide Coordinate Stability of the Latent Transforming Growth Factor-?1 Complex *"

    Article Title: Two Distinct Regions of Latency-associated Peptide Coordinate Stability of the Latent Transforming Growth Factor-?1 Complex *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.110288

    Formation of the large latent TGF-β1 complex. TGF-β1 is synthesized from a larger precursor protein, comprising a signal peptide ( SP ), LAP, and a C-terminal mature domain ( A ). During assembly, two TGF-β1 precursors are covalently
    Figure Legend Snippet: Formation of the large latent TGF-β1 complex. TGF-β1 is synthesized from a larger precursor protein, comprising a signal peptide ( SP ), LAP, and a C-terminal mature domain ( A ). During assembly, two TGF-β1 precursors are covalently

    Techniques Used: Synthesized

    Amino acids involved in the formation of the large latent TGF-β1 complex. Target residues in TGF-β1 LAP ( boldface type , underlined ) were substituted with alanine using in vitro mutagenesis (the hydrophobic residues identified to be important
    Figure Legend Snippet: Amino acids involved in the formation of the large latent TGF-β1 complex. Target residues in TGF-β1 LAP ( boldface type , underlined ) were substituted with alanine using in vitro mutagenesis (the hydrophobic residues identified to be important

    Techniques Used: In Vitro, Mutagenesis

    Analysis of the interaction between LAP and mature TGF-β1. Wild type and mutant LAP variants ( boldface type and underlined ) were generated by site-directed mutagenesis ( A ), and relative expression levels in transfected HEK-293T cells were determined
    Figure Legend Snippet: Analysis of the interaction between LAP and mature TGF-β1. Wild type and mutant LAP variants ( boldface type and underlined ) were generated by site-directed mutagenesis ( A ), and relative expression levels in transfected HEK-293T cells were determined

    Techniques Used: Mutagenesis, Generated, Expressing, Transfection

    Hydrophobic Residues at the N Terminus of LAP Govern Formation of the Small Latent TGF-β1 Complex
    Figure Legend Snippet: Hydrophobic Residues at the N Terminus of LAP Govern Formation of the Small Latent TGF-β1 Complex

    Techniques Used:

    Effects of LAP mutations on TGF-β1 biosynthesis. Target residues in TGF-β1 LAP were substituted with alanine using in vitro mutagenesis ( A ). To determine the effects of amino acid substitutions on TGF-β1 production, conditioned
    Figure Legend Snippet: Effects of LAP mutations on TGF-β1 biosynthesis. Target residues in TGF-β1 LAP were substituted with alanine using in vitro mutagenesis ( A ). To determine the effects of amino acid substitutions on TGF-β1 production, conditioned

    Techniques Used: In Vitro, Mutagenesis

    Analysis of the effect of CED mutations on TGF-β1 biosynthesis and latent complex stability. Conditioned medium from cells transfected with wild type ( WT ) or CED variants of TGF-β1 was initially analyzed by Western blot using a TGF-β1
    Figure Legend Snippet: Analysis of the effect of CED mutations on TGF-β1 biosynthesis and latent complex stability. Conditioned medium from cells transfected with wild type ( WT ) or CED variants of TGF-β1 was initially analyzed by Western blot using a TGF-β1

    Techniques Used: Transfection, Western Blot

    LAP Residues Distant from the N-terminal α-Helix Affect Stability of the Latent TGF-β1 Complex
    Figure Legend Snippet: LAP Residues Distant from the N-terminal α-Helix Affect Stability of the Latent TGF-β1 Complex

    Techniques Used:

    6) Product Images from "In vivo inhibition of rat stellate cell activation by soluble transforming growth factor ? type II receptor: A potential new therapy for hepatic fibrosis"

    Article Title: In vivo inhibition of rat stellate cell activation by soluble transforming growth factor ? type II receptor: A potential new therapy for hepatic fibrosis

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    Efficacy of sTGF-βR in blocking the actions of TGF-β. ( A ) Exogenous TGF-β produces a dose-dependent blockade of mink lung epithelial cell proliferation. ( B ) Dose-dependent inhibition of TGF-β (250 pg/ml) by soluble TGF-β receptor, measured as release of cells from the inhibitory effect of the cytokine. Serial dilutions of the sTGF-βR were incubated with a known amount of TGF-β1 (250 pg/ml) and added to the wells. Thymidine incorporation was determined over 4 hr.
    Figure Legend Snippet: Efficacy of sTGF-βR in blocking the actions of TGF-β. ( A ) Exogenous TGF-β produces a dose-dependent blockade of mink lung epithelial cell proliferation. ( B ) Dose-dependent inhibition of TGF-β (250 pg/ml) by soluble TGF-β receptor, measured as release of cells from the inhibitory effect of the cytokine. Serial dilutions of the sTGF-βR were incubated with a known amount of TGF-β1 (250 pg/ml) and added to the wells. Thymidine incorporation was determined over 4 hr.

    Techniques Used: Blocking Assay, Inhibition, Incubation

    7) Product Images from "Gelatinase A (MMP-2) Is Necessary and Sufficient for Renal Tubular Cell Epithelial-Mesenchymal Transformation"

    Article Title: Gelatinase A (MMP-2) Is Necessary and Sufficient for Renal Tubular Cell Epithelial-Mesenchymal Transformation

    Journal: The American Journal of Pathology

    doi:

    TGF-β1 induces the myofibroblastic phenotype with concurrent expression of MT1-MMP and active gelatinase A. Cultured NRK-52e cells were incubated in the presence ( panels B–H ) or absence (A) of 2 ng/ml TGF-β1 for 3 days followed by immunohistochemical analysis. A: Control cells stained for α-SMA. B: TGF-β1-treated cells stained for α-SMA showing intense expression at the migratory front. C and D: Dual immunohistochemical staining for α-SMA ( C ) and MT1-MMP ( D ) of TGF-β1-treated cells showing co localization of expression in the migratory front. E and F: Dual immunohistochemical staining for MT1-MMP ( E ) and active gelatinase A ( F ) of TGF-β1-treated cells showing co localization of enzyme proteins. G and H: Normarski optics ( G ) of TGF-β1-treated cells demonstrating migratory phenotype with acquisition of myofibroblastic morphology. H: Same cells demonstrating localization of active gelatinase A protein on the leading and trailing edges of the cells. (Magnification, ×500).
    Figure Legend Snippet: TGF-β1 induces the myofibroblastic phenotype with concurrent expression of MT1-MMP and active gelatinase A. Cultured NRK-52e cells were incubated in the presence ( panels B–H ) or absence (A) of 2 ng/ml TGF-β1 for 3 days followed by immunohistochemical analysis. A: Control cells stained for α-SMA. B: TGF-β1-treated cells stained for α-SMA showing intense expression at the migratory front. C and D: Dual immunohistochemical staining for α-SMA ( C ) and MT1-MMP ( D ) of TGF-β1-treated cells showing co localization of expression in the migratory front. E and F: Dual immunohistochemical staining for MT1-MMP ( E ) and active gelatinase A ( F ) of TGF-β1-treated cells showing co localization of enzyme proteins. G and H: Normarski optics ( G ) of TGF-β1-treated cells demonstrating migratory phenotype with acquisition of myofibroblastic morphology. H: Same cells demonstrating localization of active gelatinase A protein on the leading and trailing edges of the cells. (Magnification, ×500).

    Techniques Used: Expressing, Cell Culture, Incubation, Immunohistochemistry, Staining

    A specific gelatinase A inhibitor blocks TGF-β1-mediated NRK-52e transformation. NRK-52e cells were treated with 2 ng/ml TGF-β1 in the presence of concentrations of the cyclic peptide inhibitor, CTTHWGFTLCGG, ranging from 0, 10, 25, 50, and 100 μmol/L ( A–E , respectively). Cells in F were treated with 100 μmol/L of the control cyclic peptide, CRAVRALWRCGG. Expression of α-SMA is significantly inhibited by concentrations of the cyclic peptide gelatinase A inhibitor above 25 μmol/L, while the control peptide had no significant effect on α-SMA expression. (Magnification, ×500).
    Figure Legend Snippet: A specific gelatinase A inhibitor blocks TGF-β1-mediated NRK-52e transformation. NRK-52e cells were treated with 2 ng/ml TGF-β1 in the presence of concentrations of the cyclic peptide inhibitor, CTTHWGFTLCGG, ranging from 0, 10, 25, 50, and 100 μmol/L ( A–E , respectively). Cells in F were treated with 100 μmol/L of the control cyclic peptide, CRAVRALWRCGG. Expression of α-SMA is significantly inhibited by concentrations of the cyclic peptide gelatinase A inhibitor above 25 μmol/L, while the control peptide had no significant effect on α-SMA expression. (Magnification, ×500).

    Techniques Used: Transformation Assay, Expressing

    TGF-β1 induces MT1-MMP and gelatinase A synthesis and transcription. NRK-52e cells were treated with 2 ng/ml TGF-β1 for 3 days followed by quantitative analysis of MT1-MMP and gelatinase A protein synthesis and transcription as detailed in Materials and Methods. A: Quantitative zymography of controls and TGF-β1-treated cells demonstrated an approximate threefold increase in both MT1-MMP and gelatinase A protein synthesis. B: TGF-β1 induces an approximate twofold increase in gelatinase A transcriptional activity as measured with the promoter-luciferase construct, pT4-Luc1686, and a nearly fivefold increase in MT1-MMP transcriptional activity using the promoter-luciferase construct pMT1-Luc3280.
    Figure Legend Snippet: TGF-β1 induces MT1-MMP and gelatinase A synthesis and transcription. NRK-52e cells were treated with 2 ng/ml TGF-β1 for 3 days followed by quantitative analysis of MT1-MMP and gelatinase A protein synthesis and transcription as detailed in Materials and Methods. A: Quantitative zymography of controls and TGF-β1-treated cells demonstrated an approximate threefold increase in both MT1-MMP and gelatinase A protein synthesis. B: TGF-β1 induces an approximate twofold increase in gelatinase A transcriptional activity as measured with the promoter-luciferase construct, pT4-Luc1686, and a nearly fivefold increase in MT1-MMP transcriptional activity using the promoter-luciferase construct pMT1-Luc3280.

    Techniques Used: Zymography, Activity Assay, Luciferase, Construct

    Cell-cell contact regulates TGF-β1-mediated induction of MT1-MMP. NRK-52e cells were cultured for 3 days with TGF-β1 (2 ng/ml) followed by dual immunohistochemical staining for E-cadherin ( A ) and MT1-MMP ( B ). Arrows in A denote sites of cell-cell contact with cadherin complexes. (Magnification, ×700).
    Figure Legend Snippet: Cell-cell contact regulates TGF-β1-mediated induction of MT1-MMP. NRK-52e cells were cultured for 3 days with TGF-β1 (2 ng/ml) followed by dual immunohistochemical staining for E-cadherin ( A ) and MT1-MMP ( B ). Arrows in A denote sites of cell-cell contact with cadherin complexes. (Magnification, ×700).

    Techniques Used: Cell Culture, Immunohistochemistry, Staining

    Quantitative FACS analysis of α-SMA expression by NRK-52e cells. A: Cells were treated for 72 hours with control medium, 50 nmol/L latent gelatinase A or 50 nmol/L active gelatinase A. FACS analysis demonstrates a highly significant increase in α-SMA expression in the group treated with active gelatinase A, but not in the controls or in cells treated with latent gelatinase A. B: A gelatinase A concentration-dependent increase in α-SMA expression is demonstrated. C : Inclusion of 100 μmol/L of the gelatinase A cyclic peptide inhibitor blocked TGF-β1-mediated NRK-52e transdifferentiation as determined by α-SMA expression.
    Figure Legend Snippet: Quantitative FACS analysis of α-SMA expression by NRK-52e cells. A: Cells were treated for 72 hours with control medium, 50 nmol/L latent gelatinase A or 50 nmol/L active gelatinase A. FACS analysis demonstrates a highly significant increase in α-SMA expression in the group treated with active gelatinase A, but not in the controls or in cells treated with latent gelatinase A. B: A gelatinase A concentration-dependent increase in α-SMA expression is demonstrated. C : Inclusion of 100 μmol/L of the gelatinase A cyclic peptide inhibitor blocked TGF-β1-mediated NRK-52e transdifferentiation as determined by α-SMA expression.

    Techniques Used: FACS, Expressing, Concentration Assay

    Gelatinase A generates active TGF-β1 peptide. Cultured NRK-52e cells were maintained for 3 days in serum-free medium with concentrations of active gelatinase A ranging from 0 to 50 nmol/L. Levels of active TGF-β1 peptide were determined by ELISA and expressed as the mean ± 1 SD/100 μg cell layer protein. (*, P
    Figure Legend Snippet: Gelatinase A generates active TGF-β1 peptide. Cultured NRK-52e cells were maintained for 3 days in serum-free medium with concentrations of active gelatinase A ranging from 0 to 50 nmol/L. Levels of active TGF-β1 peptide were determined by ELISA and expressed as the mean ± 1 SD/100 μg cell layer protein. (*, P

    Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

    Calcium ionophore blocks TGF-β1-mediated NRK-52e transformation. Cells were treated with TGF-β1 (2 ng/ml) for 48 hours in the absence ( A ) or presence of ionomycin (100 and 500 nmol/L, B and C , respectively) followed by staining for α-SMA. (Magnification, ×600).
    Figure Legend Snippet: Calcium ionophore blocks TGF-β1-mediated NRK-52e transformation. Cells were treated with TGF-β1 (2 ng/ml) for 48 hours in the absence ( A ) or presence of ionomycin (100 and 500 nmol/L, B and C , respectively) followed by staining for α-SMA. (Magnification, ×600).

    Techniques Used: Transformation Assay, Staining

    8) Product Images from "Maternal country of origin, breast milk characteristics and potential influences on immunity in offspring"

    Article Title: Maternal country of origin, breast milk characteristics and potential influences on immunity in offspring

    Journal: Clinical and Experimental Immunology

    doi: 10.1111/j.1365-2249.2010.04275.x

    Breast milk TGF-β1 and TGF-β2 associates with IgA levels in breast milk
    Figure Legend Snippet: Breast milk TGF-β1 and TGF-β2 associates with IgA levels in breast milk

    Techniques Used:

    9) Product Images from "Gene-Gene Associations with the Susceptibility of Kawasaki Disease and Coronary Artery Lesions"

    Article Title: Gene-Gene Associations with the Susceptibility of Kawasaki Disease and Coronary Artery Lesions

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0143056

    Comparison cytokines levels between KD patients with high-risk genotypes and low-risk genotypes. KD patients possessing the high-risk (KD risk: 1) PDE2A (rs341058) and CYFIP2 (rs767007) genotypes of KD susceptibility (n = 49) presented with significantly lower plasma levels of TGF-β1 (9489 ± 1605 vs. 16133 ± 3015) compared to KD patients in the low-risk group (KD risk: 0, n = 24), with an odds ratio of 0.59 ( p = 0.036). (A) KD patients possessing the high-risk LOC100133214 (rs2517892) and IL2RA (rs3118470) genotypes of CAL formation (CAL risk: 1, n = 35) presented with significantly elevated plasma levels of IL-2 (14.1 ± 1.6 vs. 9.6 ± 1.2) compared to KD patients in the low-risk group (CAL risk: 0, n = 38), with an odds ratio of 1.47 ( p = 0.028). (B) KD patients possessing the high-risk LOC100133214 (rs2517892) and IL2RA (rs3118470) genotypes of CAL formation (CAL risk: 1, n = 35) presented with significantly elevated plasma levels of IL-6 (51.0 ± 14.3 vs. 18.4 ± 3.7) compared to KD patients in the low-risk group (CAL risk: 0, n = 38), with an odds ratio of 2.77 ( p = 0.033). (C) KD patients possessing the high-risk LOC100133214 (rs2517892) and IL2RA (rs3118470) genotypes of CAL formation (CAL risk: 1, n = 35) presented with significantly elevated plasma levels of IFN-γ (119.2 ± 15.2 vs. 81.8 ± 10.1) compared to KD patients in the low-risk group (CAL risk: 0, n = 38), with an odds ratio of 1.46 ( p = 0.041). (D).
    Figure Legend Snippet: Comparison cytokines levels between KD patients with high-risk genotypes and low-risk genotypes. KD patients possessing the high-risk (KD risk: 1) PDE2A (rs341058) and CYFIP2 (rs767007) genotypes of KD susceptibility (n = 49) presented with significantly lower plasma levels of TGF-β1 (9489 ± 1605 vs. 16133 ± 3015) compared to KD patients in the low-risk group (KD risk: 0, n = 24), with an odds ratio of 0.59 ( p = 0.036). (A) KD patients possessing the high-risk LOC100133214 (rs2517892) and IL2RA (rs3118470) genotypes of CAL formation (CAL risk: 1, n = 35) presented with significantly elevated plasma levels of IL-2 (14.1 ± 1.6 vs. 9.6 ± 1.2) compared to KD patients in the low-risk group (CAL risk: 0, n = 38), with an odds ratio of 1.47 ( p = 0.028). (B) KD patients possessing the high-risk LOC100133214 (rs2517892) and IL2RA (rs3118470) genotypes of CAL formation (CAL risk: 1, n = 35) presented with significantly elevated plasma levels of IL-6 (51.0 ± 14.3 vs. 18.4 ± 3.7) compared to KD patients in the low-risk group (CAL risk: 0, n = 38), with an odds ratio of 2.77 ( p = 0.033). (C) KD patients possessing the high-risk LOC100133214 (rs2517892) and IL2RA (rs3118470) genotypes of CAL formation (CAL risk: 1, n = 35) presented with significantly elevated plasma levels of IFN-γ (119.2 ± 15.2 vs. 81.8 ± 10.1) compared to KD patients in the low-risk group (CAL risk: 0, n = 38), with an odds ratio of 1.46 ( p = 0.041). (D).

    Techniques Used:

    10) Product Images from "Reversible Modulation of Myofibroblast Differentiation in Adipose-Derived Mesenchymal Stem Cells"

    Article Title: Reversible Modulation of Myofibroblast Differentiation in Adipose-Derived Mesenchymal Stem Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086865

    Differences in cell morphology with TGF-β and bFGF. ADSCs in SSFM were treated with 10/mL bFGF plus 5 µg/ml heparin (bFGF), 1 ng/mL TGF-β1 (TGFβ), or no additions (Unt). Phase contrast images were taken at 2 days (2d) and 4 days (4d) of treatment. Images are representative of more than three independent experiments. Scale bar = 50 µm.
    Figure Legend Snippet: Differences in cell morphology with TGF-β and bFGF. ADSCs in SSFM were treated with 10/mL bFGF plus 5 µg/ml heparin (bFGF), 1 ng/mL TGF-β1 (TGFβ), or no additions (Unt). Phase contrast images were taken at 2 days (2d) and 4 days (4d) of treatment. Images are representative of more than three independent experiments. Scale bar = 50 µm.

    Techniques Used:

    11) Product Images from "Loss of WISP2/CCN5 in Estrogen-Dependent MCF7 Human Breast Cancer Cells Promotes a Stem-Like Cell Phenotype"

    Article Title: Loss of WISP2/CCN5 in Estrogen-Dependent MCF7 Human Breast Cancer Cells Promotes a Stem-Like Cell Phenotype

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0087878

    WISP2 knock-down is accompanied by enhanced TGF-β signaling. ( A ) Quantification by sandwich ELISA of total (active plus latent) TGF-β1 in conditioned medium from MCF7, MCF7-sh-scrambled, MCF7-sh-WISP2 and MDAMB231 cells. Results are the means ± SD of triplicate experiments. ( B ) HEK293T cells were transfected with 0.3 µg of CAGA 9 -Luc and 10 ng of Tk-renilla as an internal control. Cells were treated with various concentrations of TGF-β (0 to 1 ng/ml) or conditioned medium (CM) from MCF7-sh-scrambled, MCF7-sh-WISP2 and MDAMB231 cells treated or not with SB43152 (10 −6 M), a specific inhibitor of TGF-β receptor kinase, for 16 h. The fold induction was determined relative to the activity of the reporter alone and represents three independent experiments assayed in triplicate. ( C ) Cells were treated with or without TGF-β (2 ng/ml) for 1 h, in the presence or absence of SB43152 (10 −6 M), for 24 h. The protein levels of Smad2, Smad3, Smad4, p-Smad2 and p-Smad3 were examined by Western blotting using the corresponding antibodies. Actin was used as a loading control. ( D ) Immunofluorescence analysis of fibronectin expression was performed for MCF7-sh-scrambled and MCF7-sh-WISP2 cells. Cells were treated as described in (C), and were then fixed, permeabilized and immunostained with anti-fibronectin and Alexa Fluor 594-conjugated secondary antibodies, as described in Materials and Methods . Nuclear DNA was stained with DAPI. Images were obtained by microscopy at ×40. Scale bars, 10 µm. *p
    Figure Legend Snippet: WISP2 knock-down is accompanied by enhanced TGF-β signaling. ( A ) Quantification by sandwich ELISA of total (active plus latent) TGF-β1 in conditioned medium from MCF7, MCF7-sh-scrambled, MCF7-sh-WISP2 and MDAMB231 cells. Results are the means ± SD of triplicate experiments. ( B ) HEK293T cells were transfected with 0.3 µg of CAGA 9 -Luc and 10 ng of Tk-renilla as an internal control. Cells were treated with various concentrations of TGF-β (0 to 1 ng/ml) or conditioned medium (CM) from MCF7-sh-scrambled, MCF7-sh-WISP2 and MDAMB231 cells treated or not with SB43152 (10 −6 M), a specific inhibitor of TGF-β receptor kinase, for 16 h. The fold induction was determined relative to the activity of the reporter alone and represents three independent experiments assayed in triplicate. ( C ) Cells were treated with or without TGF-β (2 ng/ml) for 1 h, in the presence or absence of SB43152 (10 −6 M), for 24 h. The protein levels of Smad2, Smad3, Smad4, p-Smad2 and p-Smad3 were examined by Western blotting using the corresponding antibodies. Actin was used as a loading control. ( D ) Immunofluorescence analysis of fibronectin expression was performed for MCF7-sh-scrambled and MCF7-sh-WISP2 cells. Cells were treated as described in (C), and were then fixed, permeabilized and immunostained with anti-fibronectin and Alexa Fluor 594-conjugated secondary antibodies, as described in Materials and Methods . Nuclear DNA was stained with DAPI. Images were obtained by microscopy at ×40. Scale bars, 10 µm. *p

    Techniques Used: Sandwich ELISA, Transfection, Activity Assay, Western Blot, Immunofluorescence, Expressing, Staining, Microscopy

    12) Product Images from "A Novel TGF-β Trap Blocks Chemotherapeutics-Induced TGF-β1 Signaling and Enhances Their Anticancer Activity in Gynecological Cancers"

    Article Title: A Novel TGF-β Trap Blocks Chemotherapeutics-Induced TGF-β1 Signaling and Enhances Their Anticancer Activity in Gynecological Cancers

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    doi: 10.1158/1078-0432.CCR-17-3112

    Increased stem cell markers and population in OVCAR-3 cell line by DDP and induction of promoted migration and epithelial-mesenchymal transition (EMT) by chemotherapeutics. (A) OVCAR-3 cells were treated with TGF-β1 (80pM), DDP (1μM), HTS (100nM) and RER (40nM) alone or in combination for 4 days. Total RNA extracted from the cells was used for the measurements of CD44, CD133 and ABCG transcript levels by qRT-PCR. Relative mRNA level was obtained by normalizing the Ct value of each gene transcript with the Ct value of RPL27 transcript. Data are presented as mean±SEM. * P
    Figure Legend Snippet: Increased stem cell markers and population in OVCAR-3 cell line by DDP and induction of promoted migration and epithelial-mesenchymal transition (EMT) by chemotherapeutics. (A) OVCAR-3 cells were treated with TGF-β1 (80pM), DDP (1μM), HTS (100nM) and RER (40nM) alone or in combination for 4 days. Total RNA extracted from the cells was used for the measurements of CD44, CD133 and ABCG transcript levels by qRT-PCR. Relative mRNA level was obtained by normalizing the Ct value of each gene transcript with the Ct value of RPL27 transcript. Data are presented as mean±SEM. * P

    Techniques Used: Migration, Quantitative RT-PCR

    Stimulation of TGF-β1 production and secretion in OVCAR-3 cells by chemotherapeutics. (A) qRT-PCR analysis for TGF-β1 and TGF-β2 mRNA expression in OVCAR-3 cell line after treatment with 10 μM DDP, 10 nM PTX, 100 nM Doxo, or 250 nM CPT for 24h. (B) ELISA assay of total TGF-β1 and active TGF-β1 secreted in OVCAR-3 cell conditioned medium. OVCAR-3 cell lines were plated in 6-well plates and treated with or without the indicated drugs for 24 hours in a serum-free medium. The medium was then collected for the measurements of total and active forms of TGF-β1 with a sandwich ELISA kit from R D Systems. Data presented are mean ±SEM. * P
    Figure Legend Snippet: Stimulation of TGF-β1 production and secretion in OVCAR-3 cells by chemotherapeutics. (A) qRT-PCR analysis for TGF-β1 and TGF-β2 mRNA expression in OVCAR-3 cell line after treatment with 10 μM DDP, 10 nM PTX, 100 nM Doxo, or 250 nM CPT for 24h. (B) ELISA assay of total TGF-β1 and active TGF-β1 secreted in OVCAR-3 cell conditioned medium. OVCAR-3 cell lines were plated in 6-well plates and treated with or without the indicated drugs for 24 hours in a serum-free medium. The medium was then collected for the measurements of total and active forms of TGF-β1 with a sandwich ELISA kit from R D Systems. Data presented are mean ±SEM. * P

    Techniques Used: Quantitative RT-PCR, Expressing, Cycling Probe Technology, Enzyme-linked Immunosorbent Assay, Sandwich ELISA

    13) Product Images from "TGF- β inhibition via CRISPR promotes the long-term efficacy of CAR T cells against solid tumors"

    Article Title: TGF- β inhibition via CRISPR promotes the long-term efficacy of CAR T cells against solid tumors

    Journal: JCI Insight

    doi: 10.1172/jci.insight.133977

    TGFBR2 KO improved in vivo tumor elimination efficacy of CAR T cells in CDX models after local administration. ( A ) Mesothelin and TGF-β1 expression in the CDX model. Scale bar: 200 μm. ( B ) Schematic of the in vivo experimental design using CDX models. ( C ) Fold changes of tumor volume and ( D ) mouse body weight after intratumor (i.t.) administration of CAR T cells. ( E ) Inflammatory cell infiltration of CAR T cells into liver detected by H E staining in the CDX model. Scale bar: 200 μm. ( F ) Peripheral blood analysis of the proportion of hCD3 + cells after CAR T cell i.t. administration. ( G ) Indel frequencies of M28z-TKO cells, before and after administration, determined by TIDE analysis. ( H ) Analysis of T cell subsets in peripheral blood after CAR T cell i.t. administration. M28z-TKO , TGFBR2 -KO M28z; PB, peripheral blood; CM, central memory; EM, effector memory. Mean ± SD, n = 5. Two-way ANOVA and Tukey’s multiple comparisons test was used in C and D . Ordinary 1-way ANOVA and Tukey’s multiple comparisons test was used in F . Two-way ANOVA and Sidak’s multiple comparisons test was used in H .
    Figure Legend Snippet: TGFBR2 KO improved in vivo tumor elimination efficacy of CAR T cells in CDX models after local administration. ( A ) Mesothelin and TGF-β1 expression in the CDX model. Scale bar: 200 μm. ( B ) Schematic of the in vivo experimental design using CDX models. ( C ) Fold changes of tumor volume and ( D ) mouse body weight after intratumor (i.t.) administration of CAR T cells. ( E ) Inflammatory cell infiltration of CAR T cells into liver detected by H E staining in the CDX model. Scale bar: 200 μm. ( F ) Peripheral blood analysis of the proportion of hCD3 + cells after CAR T cell i.t. administration. ( G ) Indel frequencies of M28z-TKO cells, before and after administration, determined by TIDE analysis. ( H ) Analysis of T cell subsets in peripheral blood after CAR T cell i.t. administration. M28z-TKO , TGFBR2 -KO M28z; PB, peripheral blood; CM, central memory; EM, effector memory. Mean ± SD, n = 5. Two-way ANOVA and Tukey’s multiple comparisons test was used in C and D . Ordinary 1-way ANOVA and Tukey’s multiple comparisons test was used in F . Two-way ANOVA and Sidak’s multiple comparisons test was used in H .

    Techniques Used: In Vivo, Expressing, Staining

    TGFBR2 -KO CAR T cells have potent and persistent tumor elimination efficacy in PDX tumor 1 model. ( A ) Expression of mesothelin and TGF-β1 on PDX tumor 1. Scale bar: 200 μm. ( B ) Schematic of the in vivo experimental design using PDX models. ( C ) Fold change of tumor volume after i.t. or i.v. CAR T cell administration in PDX model 1. ( D ) The proportion of hCD3 + cells in the peripheral blood of PDX tumor 1 model on day 42 after i.t. or i.v. administration of CAR T cells. ( E ) Fold change of tumor volume after PDX tumor 1 reinoculation. ( F ) The proportion of hCD3 + and GFP + cells and ( G ) T cell subsets in the peripheral blood of PDX model 1 on day 40 after tumor reinoculation. M28z-TKO, TGFBR2 -KO M28z; i.t., intratumor; PB, peripheral blood. Mean ± SD, n = 5. Two-way ANOVA and Tukey’s multiple comparisons test were used in C , E , and F . Ordinary 1-way ANOVA and Tukey’s multiple comparisons test were used in D . Two-way ANOVA and Sidak’s multiple comparisons test were used in G .
    Figure Legend Snippet: TGFBR2 -KO CAR T cells have potent and persistent tumor elimination efficacy in PDX tumor 1 model. ( A ) Expression of mesothelin and TGF-β1 on PDX tumor 1. Scale bar: 200 μm. ( B ) Schematic of the in vivo experimental design using PDX models. ( C ) Fold change of tumor volume after i.t. or i.v. CAR T cell administration in PDX model 1. ( D ) The proportion of hCD3 + cells in the peripheral blood of PDX tumor 1 model on day 42 after i.t. or i.v. administration of CAR T cells. ( E ) Fold change of tumor volume after PDX tumor 1 reinoculation. ( F ) The proportion of hCD3 + and GFP + cells and ( G ) T cell subsets in the peripheral blood of PDX model 1 on day 40 after tumor reinoculation. M28z-TKO, TGFBR2 -KO M28z; i.t., intratumor; PB, peripheral blood. Mean ± SD, n = 5. Two-way ANOVA and Tukey’s multiple comparisons test were used in C , E , and F . Ordinary 1-way ANOVA and Tukey’s multiple comparisons test were used in D . Two-way ANOVA and Sidak’s multiple comparisons test were used in G .

    Techniques Used: Expressing, In Vivo

    TGFBR2 -KO CAR T cells have potent and persistent tumor elimination efficacy in PDX tumor 2 model. ( A ) Expression of mesothelin and TGF-β1 on PDX tumor 2. Scale bar: 200 μm. ( B ) Fold change of tumor volume after i.t. CAR T cell administration in PDX model 2. ( C ) Fold change of tumor volume after PDX tumor 2 reinoculation. ( D ) Tumor sizes and weights were detected at the end of the PDX tumor 2 reinoculation experiment. M28z-TKO, TGFBR2 -KO M28z; i.t., intratumor; PB, peripheral blood; un, undetectable. Mean ± SD, n = 5, except in D . Two-way ANOVA and Tukey’s multiple comparisons test were used in B and C . Unpaired t test was used in D .
    Figure Legend Snippet: TGFBR2 -KO CAR T cells have potent and persistent tumor elimination efficacy in PDX tumor 2 model. ( A ) Expression of mesothelin and TGF-β1 on PDX tumor 2. Scale bar: 200 μm. ( B ) Fold change of tumor volume after i.t. CAR T cell administration in PDX model 2. ( C ) Fold change of tumor volume after PDX tumor 2 reinoculation. ( D ) Tumor sizes and weights were detected at the end of the PDX tumor 2 reinoculation experiment. M28z-TKO, TGFBR2 -KO M28z; i.t., intratumor; PB, peripheral blood; un, undetectable. Mean ± SD, n = 5, except in D . Two-way ANOVA and Tukey’s multiple comparisons test were used in B and C . Unpaired t test was used in D .

    Techniques Used: Expressing

    TGF-β1 affects the expression of key genes in CD4 and CD8 CAR T cells. ( A and B ) Expression changes of key functional genes in CD4 ( A ) and CD8 ( B ) cells. ( C and D ) Expression changes of target genes in CD4 ( C ) and CD8 ( D ) cells confirmed by qPCR. 4C, CD4 ctrl; 4C + T, CD4 ctrl + TGF-β1; 8C, CD8 ctrl; 8C + T, CD8 ctrl + TGF-β1; 4T, CD4-TKO; 4T + T, CD4-TKO + TGF-β1; 8T, CD8-TKO; 8T + T, CD8-TKO + TGF-β1. Mean ± SD of 3 technical repetitions in 1 assay. Ordinary 1-way ANOVA and Tukey’s multiple comparisons test were used. The assays in C and D were repeated 2 times.
    Figure Legend Snippet: TGF-β1 affects the expression of key genes in CD4 and CD8 CAR T cells. ( A and B ) Expression changes of key functional genes in CD4 ( A ) and CD8 ( B ) cells. ( C and D ) Expression changes of target genes in CD4 ( C ) and CD8 ( D ) cells confirmed by qPCR. 4C, CD4 ctrl; 4C + T, CD4 ctrl + TGF-β1; 8C, CD8 ctrl; 8C + T, CD8 ctrl + TGF-β1; 4T, CD4-TKO; 4T + T, CD4-TKO + TGF-β1; 8T, CD8-TKO; 8T + T, CD8-TKO + TGF-β1. Mean ± SD of 3 technical repetitions in 1 assay. Ordinary 1-way ANOVA and Tukey’s multiple comparisons test were used. The assays in C and D were repeated 2 times.

    Techniques Used: Expressing, Functional Assay, Real-time Polymerase Chain Reaction

    TGF-β1 induced FOXP3 -dependent iTreg-like cell conversion of CAR T cells. ( A ) FOXP3 expression in control and M28z-TKO CAR T cells after incubation with OVCAR3 cells in the presence or absence of 5 ng/mL TGF-β1. ( B ) Proliferation suppression ability of iTreg-like cells converted from control, M28z-TKO, and M28z-FKO CAR T cells after incubation with OVCAR3 cells in the presence of 5 ng/mL TGF-β1. ( C ) FOXP3 KO partially rescues the negative effects of TGF-β1 on CAR T cell–mediated tumor lysis. M28z-TKO, TGFBR2 -KO M28z; M28z-FKO, FOXP- KO M28z. Mean ± SD of 3 technical repetitions in 1 assay. Two-way ANOVA and Tukey’s multiple comparisons test were used. The assays in A and C were repeated more than 3 times and those in B were repeated 2 times.
    Figure Legend Snippet: TGF-β1 induced FOXP3 -dependent iTreg-like cell conversion of CAR T cells. ( A ) FOXP3 expression in control and M28z-TKO CAR T cells after incubation with OVCAR3 cells in the presence or absence of 5 ng/mL TGF-β1. ( B ) Proliferation suppression ability of iTreg-like cells converted from control, M28z-TKO, and M28z-FKO CAR T cells after incubation with OVCAR3 cells in the presence of 5 ng/mL TGF-β1. ( C ) FOXP3 KO partially rescues the negative effects of TGF-β1 on CAR T cell–mediated tumor lysis. M28z-TKO, TGFBR2 -KO M28z; M28z-FKO, FOXP- KO M28z. Mean ± SD of 3 technical repetitions in 1 assay. Two-way ANOVA and Tukey’s multiple comparisons test were used. The assays in A and C were repeated more than 3 times and those in B were repeated 2 times.

    Techniques Used: Expressing, Incubation, Lysis

    TGFBR2 KO rescues CAR T cell exhaustion induced by TGF-β1. ( A ) Growth curve and ( B ) representative images of M28z and M28z-TKO CAR T cells after multiple rounds of CRL5826 stimulation in the presence of 5 ng/mL TGF-β1. Scale bar: 40 μm. ( C ) Specific tumor lysis by M28z and M28z-TKO cells after 3 rounds of CRL5826 stimulation in the presence of 5 ng/mL TGF-β1. ( D ) Immune checkpoint gene expression of M28z and M28z-TKO cells after 3 rounds CRL5826 stimulation in the presence of 5 ng/mL TGF-β1. ( E ) Proportion of PD1 + cells in M28z, M28z-TKO, M28z-PKO, and M28z-DKO cells after 3 rounds of incubation with CRL5826-PDL1 in the presence of 5 ng/mL TGF-β1. ( F ) Proliferation and ( G ) tumor lysis capability of M28z, M28z-TKO, M28z-PKO, and M28z-DKO cells during 3 rounds of incubation with CRL5826-PDL1 cells in the presence of TGF-β1. M28z-TKO, TGFBR2 -KO M28z; M28z-PKO, PD1- KO M28z; M28z-DKO, TGFBR2 and PD1 double-KO M28z. Mean ± SD of 3 technical repetitions in 1 assay. Ordinary 1-way ANOVA and Tukey’s multiple comparisons test were used in C . The assays in A – C were repeated more than 3 times and those in D – G were repeated 2 times.
    Figure Legend Snippet: TGFBR2 KO rescues CAR T cell exhaustion induced by TGF-β1. ( A ) Growth curve and ( B ) representative images of M28z and M28z-TKO CAR T cells after multiple rounds of CRL5826 stimulation in the presence of 5 ng/mL TGF-β1. Scale bar: 40 μm. ( C ) Specific tumor lysis by M28z and M28z-TKO cells after 3 rounds of CRL5826 stimulation in the presence of 5 ng/mL TGF-β1. ( D ) Immune checkpoint gene expression of M28z and M28z-TKO cells after 3 rounds CRL5826 stimulation in the presence of 5 ng/mL TGF-β1. ( E ) Proportion of PD1 + cells in M28z, M28z-TKO, M28z-PKO, and M28z-DKO cells after 3 rounds of incubation with CRL5826-PDL1 in the presence of 5 ng/mL TGF-β1. ( F ) Proliferation and ( G ) tumor lysis capability of M28z, M28z-TKO, M28z-PKO, and M28z-DKO cells during 3 rounds of incubation with CRL5826-PDL1 cells in the presence of TGF-β1. M28z-TKO, TGFBR2 -KO M28z; M28z-PKO, PD1- KO M28z; M28z-DKO, TGFBR2 and PD1 double-KO M28z. Mean ± SD of 3 technical repetitions in 1 assay. Ordinary 1-way ANOVA and Tukey’s multiple comparisons test were used in C . The assays in A – C were repeated more than 3 times and those in D – G were repeated 2 times.

    Techniques Used: Lysis, Expressing, Incubation

    TGF-β1 suppresses cytolysis of CAR T cells and their ability to release cytokines via TGF-β receptor. ( A ) Specific lysis of CRL5826 tumor cells after coculture with M28z CAR T cells at a 1:1 effector/target (E/T) ratio, in the presence of 0, 2.5, 5, and 10 ng/mL TGF-β1. ( B ) IL-2 and IFN-γ secretion after coculture with M28z CAR T cells at a 1:1 E/T ratio in the presence of 5 ng/mL TGF-β1. ( C ) M28z CAR T cell–mediated tumor lysis in the presence of 5 ng/mL TGF-β1 at 0.25:1, 0.5:1 and 1:1 E/T ratios. ( D and E ) TGFBR2 KO completely rescues the negative effects of TGF-β1 on CAR T cell-mediated tumor lysis ( D ) and ( E ) IL-2 and IFN-γ secretion. M28z-TKO, TGFBR2 KO M28z. Mean ± SD of 3 technical replications per assay. Ordinary 1-way ANOVA and Dunnett’s multiple comparisons test were used in A ; 2-way ANOVA and Sidak’s multiple comparisons test were used in C ; 2-way ANOVA and Tukey’s multiple comparisons test were used in D . The assays in A , C , and D were repeated more than 3 times and those in B and E were repeated 2 times.
    Figure Legend Snippet: TGF-β1 suppresses cytolysis of CAR T cells and their ability to release cytokines via TGF-β receptor. ( A ) Specific lysis of CRL5826 tumor cells after coculture with M28z CAR T cells at a 1:1 effector/target (E/T) ratio, in the presence of 0, 2.5, 5, and 10 ng/mL TGF-β1. ( B ) IL-2 and IFN-γ secretion after coculture with M28z CAR T cells at a 1:1 E/T ratio in the presence of 5 ng/mL TGF-β1. ( C ) M28z CAR T cell–mediated tumor lysis in the presence of 5 ng/mL TGF-β1 at 0.25:1, 0.5:1 and 1:1 E/T ratios. ( D and E ) TGFBR2 KO completely rescues the negative effects of TGF-β1 on CAR T cell-mediated tumor lysis ( D ) and ( E ) IL-2 and IFN-γ secretion. M28z-TKO, TGFBR2 KO M28z. Mean ± SD of 3 technical replications per assay. Ordinary 1-way ANOVA and Dunnett’s multiple comparisons test were used in A ; 2-way ANOVA and Sidak’s multiple comparisons test were used in C ; 2-way ANOVA and Tukey’s multiple comparisons test were used in D . The assays in A , C , and D were repeated more than 3 times and those in B and E were repeated 2 times.

    Techniques Used: Lysis

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    Article Snippet: .. Quantification of TGF-β 1 To quantify the TGF-β 1 in PFP samples, the Quantikine human TGF-β1 ELISA kit (R & D Systems) was used according to the manufacturer's protocol. .. Western Blot To analyze the presence of CD73, samples of 3 μ l of each PFP or 20 ng of CD73 contained in the PFP (based on the rhCD73 type curve) were treated with Laemmli buffer and analyzed by 10% sodium dodecyl sulfate- (SDS-) polyacrylamide gel electrophoresis.

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    Article Snippet: .. TGF-β1 and TGF-β2 were quantified using a human TGF-β ELISA kit (R & D Systems, Minneapolis, MN, USA). .. The absorbance at 450 nm was determined in a microplate reader (E max; Molecular Devices, Sunnyvale, CA, USA).

    Article Title: TGFβ/Activin/Nodal Pathway in Inhibition of Human Embryonic Stem Cell Differentiation by Mechanical Strain
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    Expressing:

    Article Title: Two Distinct Regions of Latency-associated Peptide Coordinate Stability of the Latent Transforming Growth Factor-?1 Complex *
    Article Snippet: TGF-β1 LAP was detected using LAP monoclonal antibody MAB246 (R & D Systems). .. Expression of TGF-β1 was determined using a TGF-β1 monoclonal antibody (MAB240, R & D systems), specific for the mature domain of TGF-β1. ..

    other:

    Article Title: The anti-fibrotic effects of epigallocatechin-3-gallate in bile duct-ligated cholestatic rats and human hepatic stellate LX-2 cells are mediated by the PI3K/Akt/Smad pathway
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    Protein Concentration:

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    R&D Systems human tgf β 1 quantikine kit
    TGFβ1 in the human endometrium. <t>TGF-</t> <t>β</t> 1 mRNA concentrations in endometrium from across the menstrual cycle in women with HMB (blood loss > 80 mL) and NMB (blood loss
    Human Tgf β 1 Quantikine Kit, 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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    Expressions of <t>TGF-β1</t> and IL-10 protein
    Tgf β1, 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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    R&D Systems recombinant human tgf β1
    The transforming growth factor <t>(TGF)-β</t> pathway is activated during culture of mouse ATII cells. ATII cells were freshly isolated (D0) or cultured for 7 days on tissue culture plastic. The mRNA expression levels of <t>Tgf-β1</t> , Tgf-β2
    Recombinant Human Tgf β1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Human TGF beta RIII Antibody from R D Systems is a mouse monoclonal antibody to TGF beta RIII This antibody reacts with human The Human TGF beta RIII Antibody
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    TGFβ1 in the human endometrium. TGF- β 1 mRNA concentrations in endometrium from across the menstrual cycle in women with HMB (blood loss > 80 mL) and NMB (blood loss

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Reduced Transforming Growth Factor-β Activity in the Endometrium of Women With Heavy Menstrual Bleeding

    doi: 10.1210/jc.2016-3437

    Figure Lengend Snippet: TGFβ1 in the human endometrium. TGF- β 1 mRNA concentrations in endometrium from across the menstrual cycle in women with HMB (blood loss > 80 mL) and NMB (blood loss

    Article Snippet: ELISA A TGF-β 1 ELISA was performed using a Human TGF-β 1 Quantikine Kit (DB100B; R & D Systems, Loughborough, UK), according to the manufacturer's instructions.

    Techniques:

    The regulation of TGF- β 1 by cortisol in primary human endometrial stromal cells. (A) TGF- β 1 mRNA after 24-hour treatment with vehicle, cortisol (1 μM), or cortisol (1 μM) plus a TSP-1 inhibitor (5 μM LSKL). (B) Active TGF- β 1 protein levels in experimental culture supernatants following pre-ELISA acid activation of latent TGF- β 1. (C) Active TGF- β 1 protein levels in the same culture supernatants without pre-ELISA acid activation (* P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Reduced Transforming Growth Factor-β Activity in the Endometrium of Women With Heavy Menstrual Bleeding

    doi: 10.1210/jc.2016-3437

    Figure Lengend Snippet: The regulation of TGF- β 1 by cortisol in primary human endometrial stromal cells. (A) TGF- β 1 mRNA after 24-hour treatment with vehicle, cortisol (1 μM), or cortisol (1 μM) plus a TSP-1 inhibitor (5 μM LSKL). (B) Active TGF- β 1 protein levels in experimental culture supernatants following pre-ELISA acid activation of latent TGF- β 1. (C) Active TGF- β 1 protein levels in the same culture supernatants without pre-ELISA acid activation (* P

    Article Snippet: ELISA A TGF-β 1 ELISA was performed using a Human TGF-β 1 Quantikine Kit (DB100B; R & D Systems, Loughborough, UK), according to the manufacturer's instructions.

    Techniques: Enzyme-linked Immunosorbent Assay, Activation Assay

    Immunohistochemistry for TGF- β 1 in human endometrium from the perimenstrual phase. (A) Staining of late secretory (LS) and menstrual (M) phase endometrium from women with HMB ( > 80 mL) and NMB (

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Reduced Transforming Growth Factor-β Activity in the Endometrium of Women With Heavy Menstrual Bleeding

    doi: 10.1210/jc.2016-3437

    Figure Lengend Snippet: Immunohistochemistry for TGF- β 1 in human endometrium from the perimenstrual phase. (A) Staining of late secretory (LS) and menstrual (M) phase endometrium from women with HMB ( > 80 mL) and NMB (

    Article Snippet: ELISA A TGF-β 1 ELISA was performed using a Human TGF-β 1 Quantikine Kit (DB100B; R & D Systems, Loughborough, UK), according to the manufacturer's instructions.

    Techniques: Immunohistochemistry, Staining

    Proposed role of TGF- β 1 in the human endometrium at menstruation. Red stars represent findings in women with HMB and potential impact on endometrial function.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Reduced Transforming Growth Factor-β Activity in the Endometrium of Women With Heavy Menstrual Bleeding

    doi: 10.1210/jc.2016-3437

    Figure Lengend Snippet: Proposed role of TGF- β 1 in the human endometrium at menstruation. Red stars represent findings in women with HMB and potential impact on endometrial function.

    Article Snippet: ELISA A TGF-β 1 ELISA was performed using a Human TGF-β 1 Quantikine Kit (DB100B; R & D Systems, Loughborough, UK), according to the manufacturer's instructions.

    Techniques:

    The effect of TGF- β 1 on human endometrial cell wound repair. (A) Average wound scratch closure distance (scratch distance at 0 hours minus scratch distance at 24 hours) in human primary stromal endometrial cells after treatment with vehicle, the Alk receptor inhibitor SB-431542, or 1 ng TGF- β 1. (B) Images of wound scratch in HES cells treated with 10 μg/mL SB-431542 for the following: (i) 0 hours; (ii) 24 hours and treated with 1 ng TGF- β 1; (iii) 0 hours; and (iv) 24 hours. * P

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Reduced Transforming Growth Factor-β Activity in the Endometrium of Women With Heavy Menstrual Bleeding

    doi: 10.1210/jc.2016-3437

    Figure Lengend Snippet: The effect of TGF- β 1 on human endometrial cell wound repair. (A) Average wound scratch closure distance (scratch distance at 0 hours minus scratch distance at 24 hours) in human primary stromal endometrial cells after treatment with vehicle, the Alk receptor inhibitor SB-431542, or 1 ng TGF- β 1. (B) Images of wound scratch in HES cells treated with 10 μg/mL SB-431542 for the following: (i) 0 hours; (ii) 24 hours and treated with 1 ng TGF- β 1; (iii) 0 hours; and (iv) 24 hours. * P

    Article Snippet: ELISA A TGF-β 1 ELISA was performed using a Human TGF-β 1 Quantikine Kit (DB100B; R & D Systems, Loughborough, UK), according to the manufacturer's instructions.

    Techniques:

    Expressions of TGF-β1 and IL-10 protein

    Journal: Annals of Dermatology

    Article Title: The Expressions of TGF-?1 and IL-10 in Cultured Fibroblasts after ALA-IPL Photodynamic Treatment

    doi: 10.5021/ad.2011.23.1.19

    Figure Lengend Snippet: Expressions of TGF-β1 and IL-10 protein

    Article Snippet: The TGF-β1 and IL-10 ELISA kits and the monoclonal antibodies against these molecules were obtained from R & D systems (USA).

    Techniques:

    Receptor CD36 is required for apoptotic cells induced TGF-β1 mRNA and protein expression. A, CD36 protein expression in RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) for 24 h was analyzed by Western blotting with anti-CD36 (MF3) antibody, and band density was normalized to GAPDH. B, RAWTβRII cells transfected with CD36-target siRNA or control vehicle (Ctr siRNA) for 24 h were stimulated with apoptotic or viable Jurkat cells. C, Apoptotic Jurkat cells were pretreated with annexin V for 45 min and incubated with RAWTβRII transfected with CD36-target siRNA or control siRNA (Ctr siRNA). TGF-β1 mRNA expression or secreted TGF-β1 protein was analyzed as in Figure 1. D, RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) were treated with either LPS (25 ng/ml) or PMA (50 nM). The expression of TGF-β1 mRNA was analyzed as above. Values represent means ± SD of five separate experiments. ns, no significant; *, P

    Journal: PLoS ONE

    Article Title: Induction of TGF-?1 Synthesis by Macrophages in Response to Apoptotic Cells Requires Activation of the Scavenger Receptor CD36

    doi: 10.1371/journal.pone.0072772

    Figure Lengend Snippet: Receptor CD36 is required for apoptotic cells induced TGF-β1 mRNA and protein expression. A, CD36 protein expression in RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) for 24 h was analyzed by Western blotting with anti-CD36 (MF3) antibody, and band density was normalized to GAPDH. B, RAWTβRII cells transfected with CD36-target siRNA or control vehicle (Ctr siRNA) for 24 h were stimulated with apoptotic or viable Jurkat cells. C, Apoptotic Jurkat cells were pretreated with annexin V for 45 min and incubated with RAWTβRII transfected with CD36-target siRNA or control siRNA (Ctr siRNA). TGF-β1 mRNA expression or secreted TGF-β1 protein was analyzed as in Figure 1. D, RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) were treated with either LPS (25 ng/ml) or PMA (50 nM). The expression of TGF-β1 mRNA was analyzed as above. Values represent means ± SD of five separate experiments. ns, no significant; *, P

    Article Snippet: Antibodies and reagents TGF-β1 was from R & D Systems (Minneapolis, MN).

    Techniques: Expressing, Transfection, Western Blot, Incubation

    Both Lyn kinase and ERK1/2 MAPK are required for TGF-β1 synthesis induced by activating anti-CD36 mIgA. A, RAWTβRII cells were stimulated with activating anti-CD36 mIgA (2 µg/ml) for the times indicated. Total cell lysates were immunoblotted for phospho-Lyn kinase and the band density was normalized to total Lyn kinase. B, RAWTβRII cells were pretreated with the src-family kinase inhibitor PP2 (0.001 to 100 µM) for 2 h and then stimulated with anti-CD36 mIgA (2 µg/ml). After 6 h, TGF-β1 mRNA expression was analyzed by real-time PCR and normalized to GAPDH. Total TGF-β1 in the conditioned medium was analyzed by ELISA after 18 h. C, A time course of ERK1/2 phosphorylation was assessed by Western blotting in RAWTβRII cells treated with anti-CD36 mIgA (2 µg/ml). Phospho-ERK1/2 band density was normalized to total ERK1/2. D, RAWTβRII cells were preincubated with MEK kinase inhibitor U0126 or inactive analogue U0124 for 2 h and then stimulated with anti-CD36 mIgA for 6 h to detect TGF-β1 mRNA expression or for 18 h to detect secreted TGF-β1 protein as in Figure 1. Values represent means ± SD of six separate experiments.

    Journal: PLoS ONE

    Article Title: Induction of TGF-?1 Synthesis by Macrophages in Response to Apoptotic Cells Requires Activation of the Scavenger Receptor CD36

    doi: 10.1371/journal.pone.0072772

    Figure Lengend Snippet: Both Lyn kinase and ERK1/2 MAPK are required for TGF-β1 synthesis induced by activating anti-CD36 mIgA. A, RAWTβRII cells were stimulated with activating anti-CD36 mIgA (2 µg/ml) for the times indicated. Total cell lysates were immunoblotted for phospho-Lyn kinase and the band density was normalized to total Lyn kinase. B, RAWTβRII cells were pretreated with the src-family kinase inhibitor PP2 (0.001 to 100 µM) for 2 h and then stimulated with anti-CD36 mIgA (2 µg/ml). After 6 h, TGF-β1 mRNA expression was analyzed by real-time PCR and normalized to GAPDH. Total TGF-β1 in the conditioned medium was analyzed by ELISA after 18 h. C, A time course of ERK1/2 phosphorylation was assessed by Western blotting in RAWTβRII cells treated with anti-CD36 mIgA (2 µg/ml). Phospho-ERK1/2 band density was normalized to total ERK1/2. D, RAWTβRII cells were preincubated with MEK kinase inhibitor U0126 or inactive analogue U0124 for 2 h and then stimulated with anti-CD36 mIgA for 6 h to detect TGF-β1 mRNA expression or for 18 h to detect secreted TGF-β1 protein as in Figure 1. Values represent means ± SD of six separate experiments.

    Article Snippet: Antibodies and reagents TGF-β1 was from R & D Systems (Minneapolis, MN).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot

    Apoptotic cells induce TGF-β1 production in RAWTβRII macrophages. A, RAW 264.7 macrophages were stably transfected with truncated TGF-β receptor II (RAWTβRII) or empty vector (RAWV). After 48 h, the cells were incubated with TGF-β1 (25 ng/ml) for 6 h. TGF-β1 mRNA expression which was normalized to GAPDH, was measured by real-time PCR and expressed as fold enhancement. B, Human Jurkat T cells were stimulated with UV-irradiation to induce apoptosis. Surface PS exposure was assessed by annexin V staining and cell permeability with propidium iodide as analyzed by flow cytometry (shown as a representative dot plot from five independent experiments). C, Apoptotic or viable Jurkat cells were pretreated with annexin V for 45 min, and incubated with RAWTβRII macrophages for 6 h to measure TGF-β1 mRNA expression. D, Total TGF-β1 in the conditioned medium was analyzed by ELISA after 18 h of co-culture. Values represent means ± SD of five separate experiments. ***, P

    Journal: PLoS ONE

    Article Title: Induction of TGF-?1 Synthesis by Macrophages in Response to Apoptotic Cells Requires Activation of the Scavenger Receptor CD36

    doi: 10.1371/journal.pone.0072772

    Figure Lengend Snippet: Apoptotic cells induce TGF-β1 production in RAWTβRII macrophages. A, RAW 264.7 macrophages were stably transfected with truncated TGF-β receptor II (RAWTβRII) or empty vector (RAWV). After 48 h, the cells were incubated with TGF-β1 (25 ng/ml) for 6 h. TGF-β1 mRNA expression which was normalized to GAPDH, was measured by real-time PCR and expressed as fold enhancement. B, Human Jurkat T cells were stimulated with UV-irradiation to induce apoptosis. Surface PS exposure was assessed by annexin V staining and cell permeability with propidium iodide as analyzed by flow cytometry (shown as a representative dot plot from five independent experiments). C, Apoptotic or viable Jurkat cells were pretreated with annexin V for 45 min, and incubated with RAWTβRII macrophages for 6 h to measure TGF-β1 mRNA expression. D, Total TGF-β1 in the conditioned medium was analyzed by ELISA after 18 h of co-culture. Values represent means ± SD of five separate experiments. ***, P

    Article Snippet: Antibodies and reagents TGF-β1 was from R & D Systems (Minneapolis, MN).

    Techniques: Stable Transfection, Transfection, Plasmid Preparation, Incubation, Expressing, Real-time Polymerase Chain Reaction, Irradiation, Staining, Permeability, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

    Lyn kinase and ERK1/2 MAPK act by separate pathways for activating anti-CD36 mIgA-induced TGF-β1 synthesis. A, RAWTβRII cells were pretreated with inhibitor PP2 (30 µM) for 2 h prior to stimulation with anti-CD36 mIgA (2 µg/ml) for 60 min. Phosphorylation of Lyn and ERK1/2 were analyzed by Western blotting using total cell lysates. B, RAWTβRII cells were pretreated with inhibitor U0126 or analogue U0124 (1.0 µM) for 2 h and then incubated with anti-CD36 mIgA (2 µg/ml) for 90 min. Total cell lysates were used to analyze phosphorylation of Lyn and ERK1/2. Relative values for phosphorylated kinase versus total kinase were determined by densitometry and expressed as means ± SD of five separate experiments. *** P

    Journal: PLoS ONE

    Article Title: Induction of TGF-?1 Synthesis by Macrophages in Response to Apoptotic Cells Requires Activation of the Scavenger Receptor CD36

    doi: 10.1371/journal.pone.0072772

    Figure Lengend Snippet: Lyn kinase and ERK1/2 MAPK act by separate pathways for activating anti-CD36 mIgA-induced TGF-β1 synthesis. A, RAWTβRII cells were pretreated with inhibitor PP2 (30 µM) for 2 h prior to stimulation with anti-CD36 mIgA (2 µg/ml) for 60 min. Phosphorylation of Lyn and ERK1/2 were analyzed by Western blotting using total cell lysates. B, RAWTβRII cells were pretreated with inhibitor U0126 or analogue U0124 (1.0 µM) for 2 h and then incubated with anti-CD36 mIgA (2 µg/ml) for 90 min. Total cell lysates were used to analyze phosphorylation of Lyn and ERK1/2. Relative values for phosphorylated kinase versus total kinase were determined by densitometry and expressed as means ± SD of five separate experiments. *** P

    Article Snippet: Antibodies and reagents TGF-β1 was from R & D Systems (Minneapolis, MN).

    Techniques: Activated Clotting Time Assay, Western Blot, Incubation

    Lyn kinase and ERK1/2 are involved in CD36 mediated TGF-β1 induction by apoptotic cells. A, RAWTβRII cells were cultured in the presence of viable or apoptotic Jurkat cells for the time indicated. Total cell lysates were used for analyzing phosphorylation of ERK1/2 MAPK and Lyn kinase by Western blotting. B, RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) for 24 h were incubated with apoptotic Jurkat cells for 60 or 90 min to analyze phosphorylation of Lyn kinase or ERK1/2, respectively. C, RAWTβRII cells were preincubated with PP2 (30 µM) or U0126 (1 µM) for 2h and then co-cultured with apoptotic Jurkat cells. TGF-β1 mRNA expression or secreted TGF-β1 protein was analyzed as in Figure 1. Values represent means ± SD of six separate experiments. ***, P

    Journal: PLoS ONE

    Article Title: Induction of TGF-?1 Synthesis by Macrophages in Response to Apoptotic Cells Requires Activation of the Scavenger Receptor CD36

    doi: 10.1371/journal.pone.0072772

    Figure Lengend Snippet: Lyn kinase and ERK1/2 are involved in CD36 mediated TGF-β1 induction by apoptotic cells. A, RAWTβRII cells were cultured in the presence of viable or apoptotic Jurkat cells for the time indicated. Total cell lysates were used for analyzing phosphorylation of ERK1/2 MAPK and Lyn kinase by Western blotting. B, RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) for 24 h were incubated with apoptotic Jurkat cells for 60 or 90 min to analyze phosphorylation of Lyn kinase or ERK1/2, respectively. C, RAWTβRII cells were preincubated with PP2 (30 µM) or U0126 (1 µM) for 2h and then co-cultured with apoptotic Jurkat cells. TGF-β1 mRNA expression or secreted TGF-β1 protein was analyzed as in Figure 1. Values represent means ± SD of six separate experiments. ***, P

    Article Snippet: Antibodies and reagents TGF-β1 was from R & D Systems (Minneapolis, MN).

    Techniques: Cell Culture, Western Blot, Transfection, Incubation, Expressing

    Stimulation with activating anti-CD36 mIgA induces TGF-β1 synthesis. A, RAWTβRII cells were cultured with activating anti-CD36 (JC63.1) mIgA or isotype control (2 µg/ml) for the times indicated. TGF-β1 mRNA expression or secreted TGF-β1 protein was analyzed as in Figure 1. B, RAWTβRII cells were pre-treated with the indicated concentrations of actinomycin D or cycloheximide for 1 h, before stimulation for 6 h and TGF-β1 mRNA expression was analyzed as above. C, RAWTβRII cells were cultured in the presence of PMA (50 nM) for 18 h to increase the steady state TGF-β1 mRNA level, and then the cells were incubated with actinomycin D (10 µg/ml) in the presence or absence of anti-CD36 mIgA (2 µg/ml) for the times indicated. D, RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) for 24 h were incubated with anti-CD36 mIgA (2 µg/ml) or isotype control (2 µg/ml) for 6 h or 18 h to analyze TGF-β1 mRNA expression or secreted total TGF-β1 protein respectively, as in Figure 1. Values represent means ± SD of five separate experiments. **, P

    Journal: PLoS ONE

    Article Title: Induction of TGF-?1 Synthesis by Macrophages in Response to Apoptotic Cells Requires Activation of the Scavenger Receptor CD36

    doi: 10.1371/journal.pone.0072772

    Figure Lengend Snippet: Stimulation with activating anti-CD36 mIgA induces TGF-β1 synthesis. A, RAWTβRII cells were cultured with activating anti-CD36 (JC63.1) mIgA or isotype control (2 µg/ml) for the times indicated. TGF-β1 mRNA expression or secreted TGF-β1 protein was analyzed as in Figure 1. B, RAWTβRII cells were pre-treated with the indicated concentrations of actinomycin D or cycloheximide for 1 h, before stimulation for 6 h and TGF-β1 mRNA expression was analyzed as above. C, RAWTβRII cells were cultured in the presence of PMA (50 nM) for 18 h to increase the steady state TGF-β1 mRNA level, and then the cells were incubated with actinomycin D (10 µg/ml) in the presence or absence of anti-CD36 mIgA (2 µg/ml) for the times indicated. D, RAWTβRII cells transfected with CD36-target siRNA or control siRNA (Ctr siRNA) for 24 h were incubated with anti-CD36 mIgA (2 µg/ml) or isotype control (2 µg/ml) for 6 h or 18 h to analyze TGF-β1 mRNA expression or secreted total TGF-β1 protein respectively, as in Figure 1. Values represent means ± SD of five separate experiments. **, P

    Article Snippet: Antibodies and reagents TGF-β1 was from R & D Systems (Minneapolis, MN).

    Techniques: Cell Culture, Expressing, Incubation, Transfection

    The transforming growth factor (TGF)-β pathway is activated during culture of mouse ATII cells. ATII cells were freshly isolated (D0) or cultured for 7 days on tissue culture plastic. The mRNA expression levels of Tgf-β1 , Tgf-β2

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Transdifferentiation of alveolar epithelial type II to type I cells is controlled by opposing TGF-β and BMP signaling

    doi: 10.1152/ajplung.00032.2013

    Figure Lengend Snippet: The transforming growth factor (TGF)-β pathway is activated during culture of mouse ATII cells. ATII cells were freshly isolated (D0) or cultured for 7 days on tissue culture plastic. The mRNA expression levels of Tgf-β1 , Tgf-β2

    Article Snippet: Recombinant human TGF-β1 and BMP-4 were obtained from R & D Systems (Minneapolis, MN).

    Techniques: Isolation, Cell Culture, Expressing

    TGF-β1 treatment promotes ATII transdifferentiation mainly by promoting loss of ATII phenotype. ATII cells were cultured in the absence or presence of human (h) TGF-β1 (4 ng/ml) through 7 days. A : the mRNA expression of Sftpa , Sftpb , and

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Transdifferentiation of alveolar epithelial type II to type I cells is controlled by opposing TGF-β and BMP signaling

    doi: 10.1152/ajplung.00032.2013

    Figure Lengend Snippet: TGF-β1 treatment promotes ATII transdifferentiation mainly by promoting loss of ATII phenotype. ATII cells were cultured in the absence or presence of human (h) TGF-β1 (4 ng/ml) through 7 days. A : the mRNA expression of Sftpa , Sftpb , and

    Article Snippet: Recombinant human TGF-β1 and BMP-4 were obtained from R & D Systems (Minneapolis, MN).

    Techniques: Cell Culture, Expressing

    TGF-β and BMP signaling antagonizes each other during culture. Primary ATII cells were treated with hTGF-β1 (4 ng/ml) ( A ) or hBMP-4 (200 ng/ml) ( B ). Cells treated with TGF-β1 were harvested on days 1 and 3 , whereas cells treated

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    Article Title: Transdifferentiation of alveolar epithelial type II to type I cells is controlled by opposing TGF-β and BMP signaling

    doi: 10.1152/ajplung.00032.2013

    Figure Lengend Snippet: TGF-β and BMP signaling antagonizes each other during culture. Primary ATII cells were treated with hTGF-β1 (4 ng/ml) ( A ) or hBMP-4 (200 ng/ml) ( B ). Cells treated with TGF-β1 were harvested on days 1 and 3 , whereas cells treated

    Article Snippet: Recombinant human TGF-β1 and BMP-4 were obtained from R & D Systems (Minneapolis, MN).

    Techniques: