recombinant human tgfβ1  (R&D Systems)

 
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    Name:
    Recombinant Human TGF beta 1 Protein
    Description:
    The Recombinant Human TGF beta 1 Protein from R D Systems is derived from CHO The Recombinant Human TGF beta 1 Protein has been validated for the following applications Bioactivity
    Catalog Number:
    240-B-002
    Price:
    259
    Category:
    Proteins and Enzymes
    Source:
    CHO-derived Recombinant Human TGF-beta 1 Protein
    Applications:
    Bioactivity
    Purity:
    >97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie« Blue Staining.
    Conjugate:
    Unconjugated
    Size:
    2 ug
    Buy from Supplier


    Structured Review

    R&D Systems recombinant human tgfβ1
    Recombinant Human TGF beta 1 Protein
    The Recombinant Human TGF beta 1 Protein from R D Systems is derived from CHO The Recombinant Human TGF beta 1 Protein has been validated for the following applications Bioactivity
    https://www.bioz.com/result/recombinant human tgfβ1/product/R&D Systems
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant human tgfβ1 - by Bioz Stars, 2021-05
    99/100 stars

    Images

    1) Product Images from "miR-17 inhibition enhances the formation of kidney cancer spheres with stem cell/tumor initiating cell properties"

    Article Title: miR-17 inhibition enhances the formation of kidney cancer spheres with stem cell/tumor initiating cell properties

    Journal: Oncotarget

    doi:

    TGFβ signaling is involved in RCC formation and is regulated by miR-17 (A) TGFβ1 treatment induced sphere formation in ACHN cells. Parallel miR-17 transfection interfered with sphere formation. (B) TGFβ1 treated ACHN cells show increased expression of mesenchymal markers. (C) miR-17 transfection of TGFβ1 treated ACHN cells leads to decreased expression of ZEB2 and SNAIL EMT markers. (D) Silencing of TGFBR2 by siRNA or by miR-17 tra nsfection inhibited RCC sphere formation. miR-17 transfection leads to decreased TGFBR2 expression at mRNA (E) and protein level (F). (F) Quantification of Western blot detecting TGFBR2 expression. (G) CAKI-1 cells were co-transfected with the luciferase constructs containing the 3′ UTRs of TGFBR2 , SMAD4 or p21 and miR-17. Decreased luciferase activity indicates that TGFBR2 is a possible direct target of miR-17. (H) Analysis of TCGA database revealed a significant increase of CD24, CD44 and TGFβ1 expression in ccRCC patients with poor survival.
    Figure Legend Snippet: TGFβ signaling is involved in RCC formation and is regulated by miR-17 (A) TGFβ1 treatment induced sphere formation in ACHN cells. Parallel miR-17 transfection interfered with sphere formation. (B) TGFβ1 treated ACHN cells show increased expression of mesenchymal markers. (C) miR-17 transfection of TGFβ1 treated ACHN cells leads to decreased expression of ZEB2 and SNAIL EMT markers. (D) Silencing of TGFBR2 by siRNA or by miR-17 tra nsfection inhibited RCC sphere formation. miR-17 transfection leads to decreased TGFBR2 expression at mRNA (E) and protein level (F). (F) Quantification of Western blot detecting TGFBR2 expression. (G) CAKI-1 cells were co-transfected with the luciferase constructs containing the 3′ UTRs of TGFBR2 , SMAD4 or p21 and miR-17. Decreased luciferase activity indicates that TGFBR2 is a possible direct target of miR-17. (H) Analysis of TCGA database revealed a significant increase of CD24, CD44 and TGFβ1 expression in ccRCC patients with poor survival.

    Techniques Used: Transfection, Expressing, Western Blot, Luciferase, Construct, Activity Assay

    2) Product Images from "Comparing Effects of Transforming Growth Factor β1 on Microglia From Rat and Mouse: Transcriptional Profiles and Potassium Channels"

    Article Title: Comparing Effects of Transforming Growth Factor β1 on Microglia From Rat and Mouse: Transcriptional Profiles and Potassium Channels

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2018.00115

    In neonatal rat and mouse microglia, TGFβ1 did not affect Kcnj2 mRNA expression or Kir2.1 current. Microglia were unstimulated (control) or treated with TGFβ1 for 24 h (expression data) or 30 h (whole-cell recordings). (A) mRNA expression. Data are expressed as fold-change relative to control (unstimulated) microglia (mean ± SEM of 4–6 individual cultures). (B) Inward-rectifier (Kir) current. Representative traces of total inward current in control microglia, and in TGFβ1-treated cells. Whole-cell currents were recorded in response to 500 ms-long voltage steps between −170 mV and −70 mV in 10 mV increments from a holding potential of −20 mV. (C) Scatterplot of individual cells showing the proportion of the peak current (at −140 mV) that was blocked by 20 μM ML133. (D) Peak Kir2.1 current density (pA/pF) as a function of voltage. Data are shown as mean ± SEM for the number of cells indicated. (E) Summary of peak inward current density (pA/pF) measured at −140 mV. There were no significant TGFβ1 effects at the p
    Figure Legend Snippet: In neonatal rat and mouse microglia, TGFβ1 did not affect Kcnj2 mRNA expression or Kir2.1 current. Microglia were unstimulated (control) or treated with TGFβ1 for 24 h (expression data) or 30 h (whole-cell recordings). (A) mRNA expression. Data are expressed as fold-change relative to control (unstimulated) microglia (mean ± SEM of 4–6 individual cultures). (B) Inward-rectifier (Kir) current. Representative traces of total inward current in control microglia, and in TGFβ1-treated cells. Whole-cell currents were recorded in response to 500 ms-long voltage steps between −170 mV and −70 mV in 10 mV increments from a holding potential of −20 mV. (C) Scatterplot of individual cells showing the proportion of the peak current (at −140 mV) that was blocked by 20 μM ML133. (D) Peak Kir2.1 current density (pA/pF) as a function of voltage. Data are shown as mean ± SEM for the number of cells indicated. (E) Summary of peak inward current density (pA/pF) measured at −140 mV. There were no significant TGFβ1 effects at the p

    Techniques Used: Expressing, Mass Spectrometry

    Comparing effects of TGFβ1 and IL-10 on gene expression in rat and mouse microglia. Microglia were stimulated with either cytokine for 24 h. mRNA transcript levels were determined by NanoString, expressed as fold-change relative to unstimulated control cells ( p
    Figure Legend Snippet: Comparing effects of TGFβ1 and IL-10 on gene expression in rat and mouse microglia. Microglia were stimulated with either cytokine for 24 h. mRNA transcript levels were determined by NanoString, expressed as fold-change relative to unstimulated control cells ( p

    Techniques Used: Expressing

    Expression of Kcnn4 mRNA, KCa3.1 protein and current are differently affected by TGFβ1 in neonatal rat and mouse microglia. (A) Transcript expression of the Ca 2+ -activated K + channels (mean ± SEM; 4–6 individual cultures): Kcnma1 (KCa1.1/BK/maxi-K), Kcnn3 (KCa2.3/SK3), Kcnn4 (KCa3.1/SK4) and the regulatory molecule, calmodulin ( Calm1 ). (B) Representative Western blot of KCa3.1 with both species on the same gel. Quantification resulting from normalization of each KCa3.1 band to total protein (Coomassie blue staining) in each lane. Fold changes were calculated with respect to unstimulated (control) microglia (mean ± SEM; 4–7 individual cultures for mouse, 5–6 for rat). For remaining figure, left panels represent neonatal rat microglia; right panels, neonatal mouse microglia. (C) Representative whole-cell recording showing KCa3.1 current evoked by the positive gating modulator, SKA-111 (1 μM), followed by inhibition by the blocker, 1 μM TRAM-34. From a holding potential of −90 mV, the voltage was stepped to +30 mV, followed by a ramp from −120 mV to +60 mV. (D) Examples of the time course, showing activation by the positive-gating modulator, SKA-111, and inhibition by the blocker, TRAM-34. (E) Summary of KCa3.1 current activation (i.e., the TRAM-34-sensitive component) in control cells vs. TGFβ1-treated microglia. Significant differences are shown between control and TGFβ1 treated cells (*), and between species (†). One symbol of either type indicates p
    Figure Legend Snippet: Expression of Kcnn4 mRNA, KCa3.1 protein and current are differently affected by TGFβ1 in neonatal rat and mouse microglia. (A) Transcript expression of the Ca 2+ -activated K + channels (mean ± SEM; 4–6 individual cultures): Kcnma1 (KCa1.1/BK/maxi-K), Kcnn3 (KCa2.3/SK3), Kcnn4 (KCa3.1/SK4) and the regulatory molecule, calmodulin ( Calm1 ). (B) Representative Western blot of KCa3.1 with both species on the same gel. Quantification resulting from normalization of each KCa3.1 band to total protein (Coomassie blue staining) in each lane. Fold changes were calculated with respect to unstimulated (control) microglia (mean ± SEM; 4–7 individual cultures for mouse, 5–6 for rat). For remaining figure, left panels represent neonatal rat microglia; right panels, neonatal mouse microglia. (C) Representative whole-cell recording showing KCa3.1 current evoked by the positive gating modulator, SKA-111 (1 μM), followed by inhibition by the blocker, 1 μM TRAM-34. From a holding potential of −90 mV, the voltage was stepped to +30 mV, followed by a ramp from −120 mV to +60 mV. (D) Examples of the time course, showing activation by the positive-gating modulator, SKA-111, and inhibition by the blocker, TRAM-34. (E) Summary of KCa3.1 current activation (i.e., the TRAM-34-sensitive component) in control cells vs. TGFβ1-treated microglia. Significant differences are shown between control and TGFβ1 treated cells (*), and between species (†). One symbol of either type indicates p

    Techniques Used: Expressing, Western Blot, Staining, Inhibition, Activation Assay

    Effect of TGFβ1 treatment on protein levels for exemplary molecules. (A) CD206; (B) TGFβR1; (C) IFNGR1. Microglia were unstimulated (CTL, control) or stimulated with TGFβ1 for 24 h. Representative Western blots, with both species on the same gel. For each blot, the band of interest was normalized to total protein in that lane (Coomassie blue staining), and then the fold-changes were calculated with respect to unstimulated (control) microglia (mean ± SEM; 4–7 individual cultures for mouse, 5–6 for rat). Significant differences are shown between control and TGFβ1 treated cells (*), and between species (†). One symbol of either type indicates p
    Figure Legend Snippet: Effect of TGFβ1 treatment on protein levels for exemplary molecules. (A) CD206; (B) TGFβR1; (C) IFNGR1. Microglia were unstimulated (CTL, control) or stimulated with TGFβ1 for 24 h. Representative Western blots, with both species on the same gel. For each blot, the band of interest was normalized to total protein in that lane (Coomassie blue staining), and then the fold-changes were calculated with respect to unstimulated (control) microglia (mean ± SEM; 4–7 individual cultures for mouse, 5–6 for rat). Significant differences are shown between control and TGFβ1 treated cells (*), and between species (†). One symbol of either type indicates p

    Techniques Used: CTL Assay, Western Blot, Staining

    In neonatal rat and mouse microglia, TGFβ1 increases Kcna3 mRNA expression and Kv1.3 current. (A) Transcript expression of selected Kv1-family channel members. Rat and mouse microglia were unstimulated (control) or stimulated with TGFβ1 for 24 h (expression data) or 30 h (whole-cell recordings). mRNA expression (mRNA counts/200 ng total RNA) was determined by NanoString and expressed as fold-change relative to unstimulated control cells (mean ± SEM; n = 4–6 individual cultures). (B) Representative whole-cell Kv currents in control (untreated) microglia, and in TGFβ1-treated cells. The voltage clamp protocol consisted of 1 s-long voltage steps from −80 mV to +40 mV in 20 mV increments, applied every 60 s from a holding potential of −100 mV. Then, for each cell, 5 nM AgTx-2 was perfused into the bath to record the AgTx-2-insensitive component, which was then subtracted from the total current to yield the Kv1.3 current. (C) Scatter plot of individual cells showing the proportion of the peak current (at +40 mV) that was blocked by AgTx-2. (D) Peak current density (pA/pF) as a function of voltage for the total Kv current and the AgTx-2-sensitive Kv1.3 component. Data are shown as mean ± SEM for the number of cells indicated. (E) Summary of current density (pA/pF) measured at +40 mV. For all graphs, one symbol indicates p
    Figure Legend Snippet: In neonatal rat and mouse microglia, TGFβ1 increases Kcna3 mRNA expression and Kv1.3 current. (A) Transcript expression of selected Kv1-family channel members. Rat and mouse microglia were unstimulated (control) or stimulated with TGFβ1 for 24 h (expression data) or 30 h (whole-cell recordings). mRNA expression (mRNA counts/200 ng total RNA) was determined by NanoString and expressed as fold-change relative to unstimulated control cells (mean ± SEM; n = 4–6 individual cultures). (B) Representative whole-cell Kv currents in control (untreated) microglia, and in TGFβ1-treated cells. The voltage clamp protocol consisted of 1 s-long voltage steps from −80 mV to +40 mV in 20 mV increments, applied every 60 s from a holding potential of −100 mV. Then, for each cell, 5 nM AgTx-2 was perfused into the bath to record the AgTx-2-insensitive component, which was then subtracted from the total current to yield the Kv1.3 current. (C) Scatter plot of individual cells showing the proportion of the peak current (at +40 mV) that was blocked by AgTx-2. (D) Peak current density (pA/pF) as a function of voltage for the total Kv current and the AgTx-2-sensitive Kv1.3 component. Data are shown as mean ± SEM for the number of cells indicated. (E) Summary of current density (pA/pF) measured at +40 mV. For all graphs, one symbol indicates p

    Techniques Used: Expressing

    TGFβ1 increases the Kv1.3 current in adult rat and mouse microglia. Treatments and recording conditions were as in Figure 3 . (A) Scatter plot of individual cells showing the proportion of the peak current (at +40 mV) that was blocked by AgTx-2. (B) Peak current density (pA/pF) as a function of voltage for the total Kv current and the AgTx-2-sensitive Kv1.3 component. (C) Summary of current density (pA/pF) measured at +40 mV. Data are shown as mean ± SEM for the number of cells indicated. One symbol of either type indicates p
    Figure Legend Snippet: TGFβ1 increases the Kv1.3 current in adult rat and mouse microglia. Treatments and recording conditions were as in Figure 3 . (A) Scatter plot of individual cells showing the proportion of the peak current (at +40 mV) that was blocked by AgTx-2. (B) Peak current density (pA/pF) as a function of voltage for the total Kv current and the AgTx-2-sensitive Kv1.3 component. (C) Summary of current density (pA/pF) measured at +40 mV. Data are shown as mean ± SEM for the number of cells indicated. One symbol of either type indicates p

    Techniques Used:

    In adult rat and mouse microglia, TGFβ1 had no effect on the Kir2.1 current. Treatments and recording conditions were as in Figure 5 . (A) Scatterplot of individual cells showing the proportion of the peak current (at −140 mV) that was blocked by 100 μM Ba 2+ . (B) Peak Kir2.1 current density (pA/pF) as a function of voltage (mean ± SEM for the number of cells indicated). (C) Summary of peak inward current density (pA/pF) measured at −140 mV. There were no significant TGFβ1 effects at the p
    Figure Legend Snippet: In adult rat and mouse microglia, TGFβ1 had no effect on the Kir2.1 current. Treatments and recording conditions were as in Figure 5 . (A) Scatterplot of individual cells showing the proportion of the peak current (at −140 mV) that was blocked by 100 μM Ba 2+ . (B) Peak Kir2.1 current density (pA/pF) as a function of voltage (mean ± SEM for the number of cells indicated). (C) Summary of peak inward current density (pA/pF) measured at −140 mV. There were no significant TGFβ1 effects at the p

    Techniques Used:

    Transforming growth factor β1 (TGFβ1) receptor in microglia after Intracerebral hemorrhage (ICH), and general effects of TGFβ1 on microglia in vitro . (A) Microglia/macrophages increase in the hematoma (H) after ICH and express TGFβR1. Representative confocal micrographs taken at 7 days in the damaged ipsilateral rat striatum (left) and the undamaged contralateral striatum (right). The inset is a higher magnification image of the boxed area (scale bar, 20 μm) showing microglia and infiltrating macrophages in the hematoma labeled with CD11b (OX-42 antibody; red) and TGFβR1 (green). Cell nuclei are labeled with 4’-6-diamidino-2-phenylindole (DAPI; blue). Scale bar for main images is 100 μm. (B) The morphology of isolated microglia (stained for CD11b, red, as in (A) was assessed using phalloidin to stain F-actin (green) and DAPI (blue) to label nuclei. Images are representative of results from six separate cell cultures for each species. Scale bar is 100 μm. (C) Microglia proliferation was determined 24 h after TGFβ1 treatment using the CyQUANT™ assay. Results are shown as fold-change with respect to untreated (control, CTL) microglia (blue dashed line) and expressed as mean ± SEM for 9–11 individual cell cultures. (D) Apoptosis was monitored using the TiterTACS™ colorimetric assay at 24 h after TGFβ1 treatment. Results are shown as mean absorbance at 450 nm (mean ± SEM, 8–9 individual cultures). The blue dashed line indicates values for the negative controls, and the red dashed line indicates the positive controls. Significant differences are shown between control and TGFβ1 treated cells (*). One symbol of indicates p
    Figure Legend Snippet: Transforming growth factor β1 (TGFβ1) receptor in microglia after Intracerebral hemorrhage (ICH), and general effects of TGFβ1 on microglia in vitro . (A) Microglia/macrophages increase in the hematoma (H) after ICH and express TGFβR1. Representative confocal micrographs taken at 7 days in the damaged ipsilateral rat striatum (left) and the undamaged contralateral striatum (right). The inset is a higher magnification image of the boxed area (scale bar, 20 μm) showing microglia and infiltrating macrophages in the hematoma labeled with CD11b (OX-42 antibody; red) and TGFβR1 (green). Cell nuclei are labeled with 4’-6-diamidino-2-phenylindole (DAPI; blue). Scale bar for main images is 100 μm. (B) The morphology of isolated microglia (stained for CD11b, red, as in (A) was assessed using phalloidin to stain F-actin (green) and DAPI (blue) to label nuclei. Images are representative of results from six separate cell cultures for each species. Scale bar is 100 μm. (C) Microglia proliferation was determined 24 h after TGFβ1 treatment using the CyQUANT™ assay. Results are shown as fold-change with respect to untreated (control, CTL) microglia (blue dashed line) and expressed as mean ± SEM for 9–11 individual cell cultures. (D) Apoptosis was monitored using the TiterTACS™ colorimetric assay at 24 h after TGFβ1 treatment. Results are shown as mean absorbance at 450 nm (mean ± SEM, 8–9 individual cultures). The blue dashed line indicates values for the negative controls, and the red dashed line indicates the positive controls. Significant differences are shown between control and TGFβ1 treated cells (*). One symbol of indicates p

    Techniques Used: In Vitro, Labeling, Isolation, Staining, CyQUANT Assay, CTL Assay, Colorimetric Assay

    3) Product Images from "Inhibitory morphogens and monopodial branching of the embryonic chicken lung"

    Article Title: Inhibitory morphogens and monopodial branching of the embryonic chicken lung

    Journal: Developmental dynamics : an official publication of the American Association of Anatomists

    doi: 10.1002/dvdy.23771

    Effect of disrupting the endogenous TGFβ concentration ex vivo . (A) Representative brightfield images of lungs cultured over 48 hours with recombinant TGFβ1, TβRI inhibitor, or control. Images are for the same explants tracked
    Figure Legend Snippet: Effect of disrupting the endogenous TGFβ concentration ex vivo . (A) Representative brightfield images of lungs cultured over 48 hours with recombinant TGFβ1, TβRI inhibitor, or control. Images are for the same explants tracked

    Techniques Used: Concentration Assay, Ex Vivo, Cell Culture, Recombinant

    4) Product Images from "Neuropilin-2 is upregulated in lung cancer cells during TGFβ1-induced epithelial-mesenchymal transition"

    Article Title: Neuropilin-2 is upregulated in lung cancer cells during TGFβ1-induced epithelial-mesenchymal transition

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-13-1755

    Induction and maintenance of NRP2 depends on non-canonical TGFβ signaling and involves ZEB1. (A) Left : H358 cells were treated with 10 µM SB431542 (SB) or vehicle for 60 min prior to TGFβ1 exposure for the indicated times. Right:
    Figure Legend Snippet: Induction and maintenance of NRP2 depends on non-canonical TGFβ signaling and involves ZEB1. (A) Left : H358 cells were treated with 10 µM SB431542 (SB) or vehicle for 60 min prior to TGFβ1 exposure for the indicated times. Right:

    Techniques Used:

    NRP2 knockdown restores growth to TGFβ1–inhibited xenografts and reduces invasive morphology. (A) Control and H358-Tr-TGFβ cells, infected with shNRP2-B or control shRNA lentiviruses, were injected subcutaneously into nu/nu mice
    Figure Legend Snippet: NRP2 knockdown restores growth to TGFβ1–inhibited xenografts and reduces invasive morphology. (A) Control and H358-Tr-TGFβ cells, infected with shNRP2-B or control shRNA lentiviruses, were injected subcutaneously into nu/nu mice

    Techniques Used: Infection, shRNA, Injection, Mouse Assay

    TGFβ1 upregulates NRP2 protein in NSCLC cell lines. (A) Western blot showing NRP2 levels in 4 NSCLC cell lines following 1 and 3 days of TGFβ1 (10 ng/ml) treatment. EpCAM and N-cadherin were used as epithelial and mesenchymal markers,
    Figure Legend Snippet: TGFβ1 upregulates NRP2 protein in NSCLC cell lines. (A) Western blot showing NRP2 levels in 4 NSCLC cell lines following 1 and 3 days of TGFβ1 (10 ng/ml) treatment. EpCAM and N-cadherin were used as epithelial and mesenchymal markers,

    Techniques Used: Western Blot

    5) Product Images from "The role of microRNA-3085 in chondrocyte function"

    Article Title: The role of microRNA-3085 in chondrocyte function

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-78606-6

    TGFβ induces miR-3085-3p which represses Smad signaling. Primary human articular chondrocytes were cultured in micromass culture for 48 h. ( A ) cells were then treated with TGFβ1 (4 ng/ml), or control for 24 h; miR-3085-3p was measured by qRT-PCR. ( B ) Primary human articular chondrocytes were grown in monolayer culture and transiently transfected with miR-3085-3p or a non-targeting control (NTC) for 24 h prior to stimulation with TGFβ1 (4 ng/ml), or control for 6 h; ID1 was measured by qRT-PCR. ( C – E ) SW1353 cells were transiently transfected with the ( C ) Smad2, ( D ) Smad3, ( E ) Smad4. 3′UTR subcloned into the pmirGLO vector (wild-type) or a construct with miR-3085-3p seed sites mutated (mutant) with miR-3085-3p mimic or non-targeting control (NTC) for 24 h. Firefly luciferase relative light units were normalised to Renilla relative light units to give overall relative light units. Mean + /− SEM, n = 3; Student’s t-test; * p
    Figure Legend Snippet: TGFβ induces miR-3085-3p which represses Smad signaling. Primary human articular chondrocytes were cultured in micromass culture for 48 h. ( A ) cells were then treated with TGFβ1 (4 ng/ml), or control for 24 h; miR-3085-3p was measured by qRT-PCR. ( B ) Primary human articular chondrocytes were grown in monolayer culture and transiently transfected with miR-3085-3p or a non-targeting control (NTC) for 24 h prior to stimulation with TGFβ1 (4 ng/ml), or control for 6 h; ID1 was measured by qRT-PCR. ( C – E ) SW1353 cells were transiently transfected with the ( C ) Smad2, ( D ) Smad3, ( E ) Smad4. 3′UTR subcloned into the pmirGLO vector (wild-type) or a construct with miR-3085-3p seed sites mutated (mutant) with miR-3085-3p mimic or non-targeting control (NTC) for 24 h. Firefly luciferase relative light units were normalised to Renilla relative light units to give overall relative light units. Mean + /− SEM, n = 3; Student’s t-test; * p

    Techniques Used: Cell Culture, Quantitative RT-PCR, Transfection, Plasmid Preparation, Construct, Mutagenesis, Luciferase

    6) Product Images from "Reticulon 4B (Nogo-B) facilitates hepatocyte proliferation and liver regeneration"

    Article Title: Reticulon 4B (Nogo-B) facilitates hepatocyte proliferation and liver regeneration

    Journal: Hepatology (Baltimore, Md.)

    doi: 10.1002/hep.26235

    TGFβ1 and Id1 expression are greater in Nogo-B KO mice than in WT mice one day after PHx. IL-6 and EGF expression are greater in Nogo-B KO mice two days after PHx
    Figure Legend Snippet: TGFβ1 and Id1 expression are greater in Nogo-B KO mice than in WT mice one day after PHx. IL-6 and EGF expression are greater in Nogo-B KO mice two days after PHx

    Techniques Used: Expressing, Mouse Assay

    Loss of Nogo-B may sensitize hepatocytes to the inhibitory effect of TGFβ1 on proliferation
    Figure Legend Snippet: Loss of Nogo-B may sensitize hepatocytes to the inhibitory effect of TGFβ1 on proliferation

    Techniques Used:

    7) Product Images from "Concentration-dependent effects of transforming growth factor ?1 on corneal wound healing"

    Article Title: Concentration-dependent effects of transforming growth factor ?1 on corneal wound healing

    Journal: Molecular Vision

    doi:

    Increasing TGFβ1 concentrations reduces HCF migration. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and incubated for 24 h in ( A ) SSFM, ( B ) 0.01 ng/ml TGFβ1, ( C ) 0.1 ng/ml TGFβ1, ( D ) 1.0 ng/ml TGFβ1, or ( E ) imaged at time zero. Bar=200 μm. F : Using T-Scratch software, percent cell migration into the wound margin at 24 h compared to time zero was calculated. Each condition was compared to SSFM, *p-value
    Figure Legend Snippet: Increasing TGFβ1 concentrations reduces HCF migration. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and incubated for 24 h in ( A ) SSFM, ( B ) 0.01 ng/ml TGFβ1, ( C ) 0.1 ng/ml TGFβ1, ( D ) 1.0 ng/ml TGFβ1, or ( E ) imaged at time zero. Bar=200 μm. F : Using T-Scratch software, percent cell migration into the wound margin at 24 h compared to time zero was calculated. Each condition was compared to SSFM, *p-value

    Techniques Used: Migration, Incubation, Software

    Endogenously secreted TGFβ1 promotes wound healing. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and imaged ( A ) time zero, or incubated for 24 h with either ( B ) SSFM ( C ) 2 μg/ml anti-TGFβ1 antibody or ( D ) 2 μg/ml matched IgG control. Bar=200 μm, *inside lines denoting wounded area. E : Using T-Scratch software, percent cell migration into the wound margin at 24 h compared to time zero was calculated. Each condition was compared to SSFM, **p-value
    Figure Legend Snippet: Endogenously secreted TGFβ1 promotes wound healing. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and imaged ( A ) time zero, or incubated for 24 h with either ( B ) SSFM ( C ) 2 μg/ml anti-TGFβ1 antibody or ( D ) 2 μg/ml matched IgG control. Bar=200 μm, *inside lines denoting wounded area. E : Using T-Scratch software, percent cell migration into the wound margin at 24 h compared to time zero was calculated. Each condition was compared to SSFM, **p-value

    Techniques Used: Incubation, Software, Migration

    Increasing TGFβ1 concentration promotes fibrotic markers. HCFs were seeded on collagen in SSFM at 1×10 4 cells/ml on coverslips in a 24 well dish in ( A ) SSFM, ( B ) 0.01 ng/ml TGFβ1, ( C ) 0.1 ng/ml TGFβ1, or ( D ) 1.0 ng/ml TGFβ1. After 72 h ( A - D ) were immunostained for α-SMA. Bar=50 μm. For each condition, cells containing organized α-SMA stress fibers were counted ( E ) and using MetaMorph Analysis the relative cell area was quantified ( F ). For analysis greater than 100 cells per experiment were analyzed. Each condition was compared to SSFM, **p-value
    Figure Legend Snippet: Increasing TGFβ1 concentration promotes fibrotic markers. HCFs were seeded on collagen in SSFM at 1×10 4 cells/ml on coverslips in a 24 well dish in ( A ) SSFM, ( B ) 0.01 ng/ml TGFβ1, ( C ) 0.1 ng/ml TGFβ1, or ( D ) 1.0 ng/ml TGFβ1. After 72 h ( A - D ) were immunostained for α-SMA. Bar=50 μm. For each condition, cells containing organized α-SMA stress fibers were counted ( E ) and using MetaMorph Analysis the relative cell area was quantified ( F ). For analysis greater than 100 cells per experiment were analyzed. Each condition was compared to SSFM, **p-value

    Techniques Used: Concentration Assay

    Increasing TGFβ1 concentrations results in a loss of nuclear p38MAPK. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and incubated in ( A , E ) SSFM, ( B , F ) 0.01 ng/ml TGFβ1, ( C , G ) 0.1 ng/ml TGFβ1, or ( D , H ) 1.0 ng/ml TGFβ. After 4 h cells were fixed and immunostained for p38MAPK ( A - D ) or SMAD 2/3 ( E - H ). Bar=50 μm. Arrows point to nuclei in which we detected either the nuclear localization or nuclear exclusion of p38MAPK and SMAD 2/3. The percent of cells in which p38MAPK was excluded from nuclei of the leading edge cells is shown in ( I ). The percent of cells with SMAD 2/3 concentrated in nuclei in the leading edge cells is shown in ( J ). Image J software was used for quantification. Each condition was compared to SSFM. ***p-value
    Figure Legend Snippet: Increasing TGFβ1 concentrations results in a loss of nuclear p38MAPK. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and incubated in ( A , E ) SSFM, ( B , F ) 0.01 ng/ml TGFβ1, ( C , G ) 0.1 ng/ml TGFβ1, or ( D , H ) 1.0 ng/ml TGFβ. After 4 h cells were fixed and immunostained for p38MAPK ( A - D ) or SMAD 2/3 ( E - H ). Bar=50 μm. Arrows point to nuclei in which we detected either the nuclear localization or nuclear exclusion of p38MAPK and SMAD 2/3. The percent of cells in which p38MAPK was excluded from nuclei of the leading edge cells is shown in ( I ). The percent of cells with SMAD 2/3 concentrated in nuclei in the leading edge cells is shown in ( J ). Image J software was used for quantification. Each condition was compared to SSFM. ***p-value

    Techniques Used: Incubation, Software

    Nuclear p38MAPK localization is necessary for HCF migration. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and incubated in ( A , F , K ) p38MAPK inhibitor, 10 μM SB202190, ( B , G , L ) DMSO, ( C , H , M ) TGFβ1 antibody, ( D , I , N ) Control IgG, or ( E , J , O ) TGFβ RI (ALK5) inhibitor 10 μM SB431542. After 4 h cells were fixed and immunostained for p38MAPK ( A - E ) or SMAD 2/3 ( F - J ). Arrows denote the nuclei of leading edge cells in which p38MAPK and SMAD 2/3 were either localized or excluded (Bar=50 μm) or after 18 h cells were imaged ( K - O ). Bar=200 μm. DMSO is the control for addition of SB202190 or SB431542. Quantification of all data are shown in the bar graphs below the images. Left to right: Exclusion of p38MAPK from the nucleus in leading edge cells, exclusion of SMAD 2/3 from the nucleus in leading edge cells, cell migration into the wound. Nuclear localization was counted using Image J software. Two non-biased people scored cell migration, 0 (less migration) to 5 (most migration). **p-value
    Figure Legend Snippet: Nuclear p38MAPK localization is necessary for HCF migration. HCFs were seeded on collagen in SSFM at 1×10 5 cells/ml in a 24 well dish. The next day cells were scratch-wounded and incubated in ( A , F , K ) p38MAPK inhibitor, 10 μM SB202190, ( B , G , L ) DMSO, ( C , H , M ) TGFβ1 antibody, ( D , I , N ) Control IgG, or ( E , J , O ) TGFβ RI (ALK5) inhibitor 10 μM SB431542. After 4 h cells were fixed and immunostained for p38MAPK ( A - E ) or SMAD 2/3 ( F - J ). Arrows denote the nuclei of leading edge cells in which p38MAPK and SMAD 2/3 were either localized or excluded (Bar=50 μm) or after 18 h cells were imaged ( K - O ). Bar=200 μm. DMSO is the control for addition of SB202190 or SB431542. Quantification of all data are shown in the bar graphs below the images. Left to right: Exclusion of p38MAPK from the nucleus in leading edge cells, exclusion of SMAD 2/3 from the nucleus in leading edge cells, cell migration into the wound. Nuclear localization was counted using Image J software. Two non-biased people scored cell migration, 0 (less migration) to 5 (most migration). **p-value

    Techniques Used: Migration, Incubation, Software

    8) Product Images from "Omega-3 Polyunsaturated Fatty Acids Attenuate Fibroblast Activation and Kidney Fibrosis Involving MTORC2 Signaling Suppression"

    Article Title: Omega-3 Polyunsaturated Fatty Acids Attenuate Fibroblast Activation and Kidney Fibrosis Involving MTORC2 Signaling Suppression

    Journal: Scientific Reports

    doi: 10.1038/srep46146

    DHA inhibits TGFβ1-stimulated Akt phosphorylation in NRK-49F cells. Cultured NRK-49F cells were serum-starved for 24 h, then administrated with 20 uM DHA for 30 min, followed by TGFβ1 (2 ng/ml) treatment for various time points as indicated. ( A ) Western blotting analyses showing that DHA treatment could decrease the abundance of p-Akt (Ser473) and p-Akt (Thr308) but not p-S6 or p-Smad3 induced by TGFβ1 in NRK-49F cells. ( B ) Quantitative analysis for p-Akt (Ser473), p-Akt (Thr308), p-S6 and p-Smad3. *p
    Figure Legend Snippet: DHA inhibits TGFβ1-stimulated Akt phosphorylation in NRK-49F cells. Cultured NRK-49F cells were serum-starved for 24 h, then administrated with 20 uM DHA for 30 min, followed by TGFβ1 (2 ng/ml) treatment for various time points as indicated. ( A ) Western blotting analyses showing that DHA treatment could decrease the abundance of p-Akt (Ser473) and p-Akt (Thr308) but not p-S6 or p-Smad3 induced by TGFβ1 in NRK-49F cells. ( B ) Quantitative analysis for p-Akt (Ser473), p-Akt (Thr308), p-S6 and p-Smad3. *p

    Techniques Used: Cell Culture, Western Blot

    DHA inhibits fibroblast activation induced by TGFβ1. ( A ) Western blotting analysis showing that DHA could time-dependently inhibit the expression of FN, α-SMA and type I collagen stimulated by TGFβ1 treatment. ( B ) Western blotting analysis showing that DHA could dose-dependently inhibit TGFβ1-induced FN, type I collagen and α-SMA expression. ( C ) Representative micrographs showing the immune-staining for FN and α-SMA in NRK-49F cells.
    Figure Legend Snippet: DHA inhibits fibroblast activation induced by TGFβ1. ( A ) Western blotting analysis showing that DHA could time-dependently inhibit the expression of FN, α-SMA and type I collagen stimulated by TGFβ1 treatment. ( B ) Western blotting analysis showing that DHA could dose-dependently inhibit TGFβ1-induced FN, type I collagen and α-SMA expression. ( C ) Representative micrographs showing the immune-staining for FN and α-SMA in NRK-49F cells.

    Techniques Used: Activation Assay, Western Blot, Expressing, Staining

    9) Product Images from "Potent Anti-Inflammatory and Neuroprotective Effects of TGF?1 Are Mediated through the Inhibition of ERK and p47phox-Ser345 Phosphorylation and Translocation in Microglia"

    Article Title: Potent Anti-Inflammatory and Neuroprotective Effects of TGF?1 Are Mediated through the Inhibition of ERK and p47phox-Ser345 Phosphorylation and Translocation in Microglia

    Journal:

    doi:

    TGFβ1 protects DA neurons against MPP + -induced toxicity in neuron-glia cultures. Different doses of TGFβl were added to the neuron-glia cultures ( A ), or neuron-enriched cultures ( B: open bars ) and neuron-astrocyte cultures ( B: closed bars
    Figure Legend Snippet: TGFβ1 protects DA neurons against MPP + -induced toxicity in neuron-glia cultures. Different doses of TGFβl were added to the neuron-glia cultures ( A ), or neuron-enriched cultures ( B: open bars ) and neuron-astrocyte cultures ( B: closed bars

    Techniques Used:

    Effects of TGFβ1 on cytosolic p47 phox protein translocation. ( A ): HAPI cells seeded in dish at 5 × 10 4 cells/well were treated with LPS for 10 min in the absence or presence of TGFβ1 pretreatment for 1 h. Cells were fixed with 3.7%
    Figure Legend Snippet: Effects of TGFβ1 on cytosolic p47 phox protein translocation. ( A ): HAPI cells seeded in dish at 5 × 10 4 cells/well were treated with LPS for 10 min in the absence or presence of TGFβ1 pretreatment for 1 h. Cells were fixed with 3.7%

    Techniques Used: Translocation Assay

    Microglia NADPH oxidase is the target of TGFβ1 inhibition in LPS-induced neurotoxicity. PHOX +/+ and PHOX –/– mice neuron-glia cultures were pretreated with vehicle or TGFβ1 for 1 h followed by LPS treatment. Neurotoxicity
    Figure Legend Snippet: Microglia NADPH oxidase is the target of TGFβ1 inhibition in LPS-induced neurotoxicity. PHOX +/+ and PHOX –/– mice neuron-glia cultures were pretreated with vehicle or TGFβ1 for 1 h followed by LPS treatment. Neurotoxicity

    Techniques Used: Inhibition, Mouse Assay

    Western blot analysis of p47 phox and ERK phosphorylation. Enriched microglial cells were treated with LPS (10 ng/ml) in the presence or absence of TGFβ1 (3 ng/ml) for 10 min, cells were then harvested, proteins were analyzed by SDS-PAGE and immunoblotting
    Figure Legend Snippet: Western blot analysis of p47 phox and ERK phosphorylation. Enriched microglial cells were treated with LPS (10 ng/ml) in the presence or absence of TGFβ1 (3 ng/ml) for 10 min, cells were then harvested, proteins were analyzed by SDS-PAGE and immunoblotting

    Techniques Used: Western Blot, SDS Page

    TGFβ1 protects DA neurons against LPS-induced toxicity. Rat primary mesencephalic neuron-glia cultures were seeded in a 24-well culture plate at 5×10 5 , then pretreated with various concentrations of TGFβ1 for 1 h before the addition
    Figure Legend Snippet: TGFβ1 protects DA neurons against LPS-induced toxicity. Rat primary mesencephalic neuron-glia cultures were seeded in a 24-well culture plate at 5×10 5 , then pretreated with various concentrations of TGFβ1 for 1 h before the addition

    Techniques Used:

    10) Product Images from "Small-Diameter Vascular Graft Engineered Using Human Embryonic Stem Cell-Derived Mesenchymal Cells"

    Article Title: Small-Diameter Vascular Graft Engineered Using Human Embryonic Stem Cell-Derived Mesenchymal Cells

    Journal: Tissue Engineering. Part A

    doi: 10.1089/ten.tea.2012.0738

    TGFβ1 induces the differentiation of hES-MSCs into calponin-positive smooth muscle-like cells. (A) Immunofluorescent staining for SMC markers, such as (1) αSMA and (2) calponin. (B) Western blot analysis for protein detection. Cells treated with various concentrations of (a) TGFβ1 or (b) serum over 7 to 14 days. (C) Enzyme-linked immunosorbent assay (ELISA) for TGFβ1 in various media. * p
    Figure Legend Snippet: TGFβ1 induces the differentiation of hES-MSCs into calponin-positive smooth muscle-like cells. (A) Immunofluorescent staining for SMC markers, such as (1) αSMA and (2) calponin. (B) Western blot analysis for protein detection. Cells treated with various concentrations of (a) TGFβ1 or (b) serum over 7 to 14 days. (C) Enzyme-linked immunosorbent assay (ELISA) for TGFβ1 in various media. * p

    Techniques Used: Staining, Western Blot, Enzyme-linked Immunosorbent Assay

    Functional characterization of hES-MSCs. (A, B) TGFβ1/ALK5 pathway is active in hES-MSCs. (A) Real-time PCR data (normalized to GAPDH) for various SMC genes: (a) αSMA, (b) CNN1, (c) SM22α. (B) Downregulation of αSMA in the presence of ALK5 inhibitor by immunostaining. (C, D) hES-MSCs display a contractile phenotype. (C) Gross images of seeded collagen gels. (D) Quantitation of gel surface area. * p
    Figure Legend Snippet: Functional characterization of hES-MSCs. (A, B) TGFβ1/ALK5 pathway is active in hES-MSCs. (A) Real-time PCR data (normalized to GAPDH) for various SMC genes: (a) αSMA, (b) CNN1, (c) SM22α. (B) Downregulation of αSMA in the presence of ALK5 inhibitor by immunostaining. (C, D) hES-MSCs display a contractile phenotype. (C) Gross images of seeded collagen gels. (D) Quantitation of gel surface area. * p

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

    11) Product Images from "Orbital Fibroblasts From Thyroid Eye Disease Patients Differ in Proliferative and Adipogenic Responses Depending on Disease Subtype"

    Article Title: Orbital Fibroblasts From Thyroid Eye Disease Patients Differ in Proliferative and Adipogenic Responses Depending on Disease Subtype

    Journal: Investigative Ophthalmology & Visual Science

    doi: 10.1167/iovs.13-12741

    Type II OFs proliferate more than type I OFs with 5 ng/mL TGFβ1 treatment. Orbital fibroblasts from three type I patients, three type II patients, and three non-TED patients were treated with 0.5% FBS culture media with or without 5 ng/mL TGFβ1.
    Figure Legend Snippet: Type II OFs proliferate more than type I OFs with 5 ng/mL TGFβ1 treatment. Orbital fibroblasts from three type I patients, three type II patients, and three non-TED patients were treated with 0.5% FBS culture media with or without 5 ng/mL TGFβ1.

    Techniques Used:

    12) Product Images from "The role of microRNA-3085 in chondrocyte function"

    Article Title: The role of microRNA-3085 in chondrocyte function

    Journal: Scientific Reports

    doi: 10.1038/s41598-020-78606-6

    TGFβ induces miR-3085-3p which represses Smad signaling. Primary human articular chondrocytes were cultured in micromass culture for 48 h. ( A ) cells were then treated with TGFβ1 (4 ng/ml), or control for 24 h; miR-3085-3p was measured by qRT-PCR. ( B ) Primary human articular chondrocytes were grown in monolayer culture and transiently transfected with miR-3085-3p or a non-targeting control (NTC) for 24 h prior to stimulation with TGFβ1 (4 ng/ml), or control for 6 h; ID1 was measured by qRT-PCR. ( C – E ) SW1353 cells were transiently transfected with the ( C ) Smad2, ( D ) Smad3, ( E ) Smad4. 3′UTR subcloned into the pmirGLO vector (wild-type) or a construct with miR-3085-3p seed sites mutated (mutant) with miR-3085-3p mimic or non-targeting control (NTC) for 24 h. Firefly luciferase relative light units were normalised to Renilla relative light units to give overall relative light units. Mean + /− SEM, n = 3; Student’s t-test; * p
    Figure Legend Snippet: TGFβ induces miR-3085-3p which represses Smad signaling. Primary human articular chondrocytes were cultured in micromass culture for 48 h. ( A ) cells were then treated with TGFβ1 (4 ng/ml), or control for 24 h; miR-3085-3p was measured by qRT-PCR. ( B ) Primary human articular chondrocytes were grown in monolayer culture and transiently transfected with miR-3085-3p or a non-targeting control (NTC) for 24 h prior to stimulation with TGFβ1 (4 ng/ml), or control for 6 h; ID1 was measured by qRT-PCR. ( C – E ) SW1353 cells were transiently transfected with the ( C ) Smad2, ( D ) Smad3, ( E ) Smad4. 3′UTR subcloned into the pmirGLO vector (wild-type) or a construct with miR-3085-3p seed sites mutated (mutant) with miR-3085-3p mimic or non-targeting control (NTC) for 24 h. Firefly luciferase relative light units were normalised to Renilla relative light units to give overall relative light units. Mean + /− SEM, n = 3; Student’s t-test; * p

    Techniques Used: Cell Culture, Quantitative RT-PCR, Transfection, Plasmid Preparation, Construct, Mutagenesis, Luciferase

    13) Product Images from "Targeting the Notch and TGF-β signaling pathways to prevent retinal fibrosis in vitro and in vivo"

    Article Title: Targeting the Notch and TGF-β signaling pathways to prevent retinal fibrosis in vitro and in vivo

    Journal: Theranostics

    doi: 10.7150/thno.45192

    Notch inhibition reduced expression of ECM proteins in human Müller cells treated with Notch and TGFβ ligands. RO inhibited the overexpression of fibronectin (A) and integrin α5 (B) in Müller cells treated with Notch ligands Dll4 and Jagged1 (both 50 ng/ml) and TGFβ1 (10 ng/ml) for 24 hours. ****P
    Figure Legend Snippet: Notch inhibition reduced expression of ECM proteins in human Müller cells treated with Notch and TGFβ ligands. RO inhibited the overexpression of fibronectin (A) and integrin α5 (B) in Müller cells treated with Notch ligands Dll4 and Jagged1 (both 50 ng/ml) and TGFβ1 (10 ng/ml) for 24 hours. ****P

    Techniques Used: Inhibition, Expressing, Over Expression

    Intravitreal injection of RO4929097 (RO) inhibits Notch and TGFβ signalling in mice with NaIO 3 -induced retinal damage. Mice received intravitreal injections of RO or vehicle (Ve) immediately after NaIO 3 -induced retinal damage and data analyses were conducted 5 days later. (A-D) Western blot analyses of Notch signalling proteins including Notch ligands Dll4 and Jagged1 (A) , Notch receptor 1 and γ-secretase presenilin 2 (B) , Notch downstream effectors including Hes1 and Hes5 (C) as well as TGFβ signalling proteins including TGFβ1 and p-Smad3 (D) . *P
    Figure Legend Snippet: Intravitreal injection of RO4929097 (RO) inhibits Notch and TGFβ signalling in mice with NaIO 3 -induced retinal damage. Mice received intravitreal injections of RO or vehicle (Ve) immediately after NaIO 3 -induced retinal damage and data analyses were conducted 5 days later. (A-D) Western blot analyses of Notch signalling proteins including Notch ligands Dll4 and Jagged1 (A) , Notch receptor 1 and γ-secretase presenilin 2 (B) , Notch downstream effectors including Hes1 and Hes5 (C) as well as TGFβ signalling proteins including TGFβ1 and p-Smad3 (D) . *P

    Techniques Used: Injection, Mouse Assay, Western Blot

    RO4929097 (RO) inhibits Notch and TGFβ signalling in Müller cells stimulated by Notch ligands. MIO-M1 human Müller cells were cultured in normal (control, Ctl) and test media containing Notch ligands including Dll4 and Jagged1 (both 50 ng/ml), either with or without RO (10 μM) for 18 hours. (A) Treatment of Müller cells with Dll4 and Jag1 upregulated the expression of γ-secretase proteinases including nicastrin, presenilin 1 and 2 as well as presenilin enhancer 2 (PEN2), all of which were significantly inhibited by the selective γ-secretase inhibitor RO. (B and C) Treatment of Müller cells with Dll4 and Jag1 also upregulated the expression of TGFβ1, TGFβ receptors 1 and 2 (TGFβ R1 and TGFβ R2, ( B ) and p-Smad3 ( C ), all of which were significantly inhibited by RO treatment. *P
    Figure Legend Snippet: RO4929097 (RO) inhibits Notch and TGFβ signalling in Müller cells stimulated by Notch ligands. MIO-M1 human Müller cells were cultured in normal (control, Ctl) and test media containing Notch ligands including Dll4 and Jagged1 (both 50 ng/ml), either with or without RO (10 μM) for 18 hours. (A) Treatment of Müller cells with Dll4 and Jag1 upregulated the expression of γ-secretase proteinases including nicastrin, presenilin 1 and 2 as well as presenilin enhancer 2 (PEN2), all of which were significantly inhibited by the selective γ-secretase inhibitor RO. (B and C) Treatment of Müller cells with Dll4 and Jag1 also upregulated the expression of TGFβ1, TGFβ receptors 1 and 2 (TGFβ R1 and TGFβ R2, ( B ) and p-Smad3 ( C ), all of which were significantly inhibited by RO treatment. *P

    Techniques Used: Cell Culture, Expressing

    RO4929097 (RO) inhibits TGFβ and Notch signalling in Müller cells stimulated by TGFβ1. Müller cells were cultured in normal (control, Ctl) and test media supplemented with recombinant human TGFβ1 (10 ng/ml), with or without RO (10 μM) for 18 hours. (A and B) TGFβ1 treatment upregulated expression of TGFβ receptors including TGFβ R1 and 2, endogenous TGFβ1 (A) and p-Smad3 (B) while this effect was significantly inhibited by RO. (C and D) TGFβ1 also significantly upregulated expression of γ-secretase proteinases including nicastrin, presenilin 1 and 2, presenilin enhancer 2 (PEN2) as well as Notch downstream effectors including Hes1 and Hes5, while this effect was inhibited by RO treatment. *P
    Figure Legend Snippet: RO4929097 (RO) inhibits TGFβ and Notch signalling in Müller cells stimulated by TGFβ1. Müller cells were cultured in normal (control, Ctl) and test media supplemented with recombinant human TGFβ1 (10 ng/ml), with or without RO (10 μM) for 18 hours. (A and B) TGFβ1 treatment upregulated expression of TGFβ receptors including TGFβ R1 and 2, endogenous TGFβ1 (A) and p-Smad3 (B) while this effect was significantly inhibited by RO. (C and D) TGFβ1 also significantly upregulated expression of γ-secretase proteinases including nicastrin, presenilin 1 and 2, presenilin enhancer 2 (PEN2) as well as Notch downstream effectors including Hes1 and Hes5, while this effect was inhibited by RO treatment. *P

    Techniques Used: Cell Culture, Recombinant, Expressing

    Notch and TGFβ ligands have additive effects on upregulation of extracellular matrix (ECM) proteins in human Müller cells. (A) Western blots for ECM proteins including fibronectin, intergrin α5 and matrix metallopeptidase 2 (MMP2) in Müller cells treated with Notch ligands Dll4 and Jagged 1 (Jag1, both 50 ng/ml) in combination with TGFβ1 (10 ng/ml) for 24, 48 or 72 hours. (B-D) Quantitative Western blot analyses of expression of fibronectin (B), intergrin α5 (C) and MMP2 (D) in Müller cells treated with Dll4 and Jagged1 (both 50 ng/ml), TGFβ1 (10 ng/ml) or a combination of both for 48 hours. A combination of Notch ligands and TGFβ showed additive effects on promoting overexpression of fibronectin, intergrin α5 and MMP2. *P
    Figure Legend Snippet: Notch and TGFβ ligands have additive effects on upregulation of extracellular matrix (ECM) proteins in human Müller cells. (A) Western blots for ECM proteins including fibronectin, intergrin α5 and matrix metallopeptidase 2 (MMP2) in Müller cells treated with Notch ligands Dll4 and Jagged 1 (Jag1, both 50 ng/ml) in combination with TGFβ1 (10 ng/ml) for 24, 48 or 72 hours. (B-D) Quantitative Western blot analyses of expression of fibronectin (B), intergrin α5 (C) and MMP2 (D) in Müller cells treated with Dll4 and Jagged1 (both 50 ng/ml), TGFβ1 (10 ng/ml) or a combination of both for 48 hours. A combination of Notch ligands and TGFβ showed additive effects on promoting overexpression of fibronectin, intergrin α5 and MMP2. *P

    Techniques Used: Western Blot, Expressing, Over Expression

    RO4929097 (RO) inhibits overexpression of ECM proteins in human Müller cells treated with Notch and TGFβ ligands. Treatment of Müller cells with Notch ligands (Dll4 and Jagged1, both 50 ng/ml) or TGFβ1 (10 ng/ml) for 24 hours promoted expression of fibronectin, intergrin α5 (A) and MMP2 (B) , with the most profound effects observed in the group treated with Notch ligands in combination with TGFβ1. RO treatment significantly inhibited overexpression of fibronectin, intergrin α5 and MMP2 in Müller cells stimulated by Notch ligands, TGFβ1 or a combination of both. *P
    Figure Legend Snippet: RO4929097 (RO) inhibits overexpression of ECM proteins in human Müller cells treated with Notch and TGFβ ligands. Treatment of Müller cells with Notch ligands (Dll4 and Jagged1, both 50 ng/ml) or TGFβ1 (10 ng/ml) for 24 hours promoted expression of fibronectin, intergrin α5 (A) and MMP2 (B) , with the most profound effects observed in the group treated with Notch ligands in combination with TGFβ1. RO treatment significantly inhibited overexpression of fibronectin, intergrin α5 and MMP2 in Müller cells stimulated by Notch ligands, TGFβ1 or a combination of both. *P

    Techniques Used: Over Expression, Expressing

    Optimising timepoints for activation of Notch and TGFβ signalling pathways in MIO-M1 human Müller cells. Western blots were conducted after culturing Müller cells in normal (control, Ctl) and test media containing recombinant human Notch ligands including Dll4 and Jagged 1 (Jag1, both 50 ng/ml) or TGFβ1 (10 ng/ml) for 3, 6 18 and 24 hours. (A) Changes in γ-secretase proteinases including nicastrin, presenilin 1 and 2, presenilin enhancer 2 (PEN2) and Notch downstream effectors including endogenous TGFβ1, Hes1 and Hes5. (B) Changes in total and phosphorylated Smad 2/3 (p-Smad 2/3). Treatment of Müller cells with Notch or TGFβ ligands for 18 or 24 hours induced extensive activation of both Notch and TGFβ signalling pathways. Independent repeats=2. (C-E) Densitometry measurements of proteins in the Notch and TGF signaling pathways after normalization to the housekeeping protein β-actin.
    Figure Legend Snippet: Optimising timepoints for activation of Notch and TGFβ signalling pathways in MIO-M1 human Müller cells. Western blots were conducted after culturing Müller cells in normal (control, Ctl) and test media containing recombinant human Notch ligands including Dll4 and Jagged 1 (Jag1, both 50 ng/ml) or TGFβ1 (10 ng/ml) for 3, 6 18 and 24 hours. (A) Changes in γ-secretase proteinases including nicastrin, presenilin 1 and 2, presenilin enhancer 2 (PEN2) and Notch downstream effectors including endogenous TGFβ1, Hes1 and Hes5. (B) Changes in total and phosphorylated Smad 2/3 (p-Smad 2/3). Treatment of Müller cells with Notch or TGFβ ligands for 18 or 24 hours induced extensive activation of both Notch and TGFβ signalling pathways. Independent repeats=2. (C-E) Densitometry measurements of proteins in the Notch and TGF signaling pathways after normalization to the housekeeping protein β-actin.

    Techniques Used: Activation Assay, Western Blot, Recombinant

    14) Product Images from "A switch from CD44+ cell to EMT cell drives the metastasis of prostate cancer"

    Article Title: A switch from CD44+ cell to EMT cell drives the metastasis of prostate cancer

    Journal: Oncotarget

    doi:

    TGFβ1 can activate the dedifferentiation of PCa cells, leading to increased CD44+ S/P cell population (a) TGFβ1 was detected by IHC in human PCa samples before ADT, after ADT for 3 month, and of CRPC specimens. TGFβ 1 expression was increased after ADT therapy. (b) TGFβ1 and phospho-Smad2/3 expression were increased in 16wks and 24wks castrated TRAMP prostate tumors comparing to wt TRAMP tumors. (c) TGFβ 1 could expanse CD44 + cell population in vitro . LNCaP cells and CWR22rv1 cells were treated by 5ng/ml TGFβ 1 for 3, 6, 9, and 12hrs. The CD44 + cell population was separated by flow cytometry. (d) 5ng/ml TGFβ 1 increased the expression of cancer stem-like cell markers, including CD44, Oct-4, c-met, nanog and sox2 in LNCaP cells by real-time PCR assay. Data are in triplicate from three independent experiments and were normalized to GAPDH. All data are expressed as mean±S.D. (e) 5ng/ml TGFβ 1 could also induce more sphere formation the character of S/P cells, compared to non-treatment LNCaP cells. Quantitation of the numbers of spheres (diameter > 40μm) are presented as the mean SD ( Scale bar, 100μm). (f) 5ng/ml TGFβ 1 could up-regulate the expression of CD44 in LNCaP and CWR22RV1 cells via Western blot assay.(g) Importantly, We also applied interruption approach via using SB431542 to block TGFβ 1 signaling and found CD44 expression decreased in Western blot assay. Significance was defined as p
    Figure Legend Snippet: TGFβ1 can activate the dedifferentiation of PCa cells, leading to increased CD44+ S/P cell population (a) TGFβ1 was detected by IHC in human PCa samples before ADT, after ADT for 3 month, and of CRPC specimens. TGFβ 1 expression was increased after ADT therapy. (b) TGFβ1 and phospho-Smad2/3 expression were increased in 16wks and 24wks castrated TRAMP prostate tumors comparing to wt TRAMP tumors. (c) TGFβ 1 could expanse CD44 + cell population in vitro . LNCaP cells and CWR22rv1 cells were treated by 5ng/ml TGFβ 1 for 3, 6, 9, and 12hrs. The CD44 + cell population was separated by flow cytometry. (d) 5ng/ml TGFβ 1 increased the expression of cancer stem-like cell markers, including CD44, Oct-4, c-met, nanog and sox2 in LNCaP cells by real-time PCR assay. Data are in triplicate from three independent experiments and were normalized to GAPDH. All data are expressed as mean±S.D. (e) 5ng/ml TGFβ 1 could also induce more sphere formation the character of S/P cells, compared to non-treatment LNCaP cells. Quantitation of the numbers of spheres (diameter > 40μm) are presented as the mean SD ( Scale bar, 100μm). (f) 5ng/ml TGFβ 1 could up-regulate the expression of CD44 in LNCaP and CWR22RV1 cells via Western blot assay.(g) Importantly, We also applied interruption approach via using SB431542 to block TGFβ 1 signaling and found CD44 expression decreased in Western blot assay. Significance was defined as p

    Techniques Used: Immunohistochemistry, Expressing, In Vitro, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction, Quantitation Assay, Western Blot, Blocking Assay

    TGFβ1 altered EMT with enhanced PCa invasion via modulating CD44 (a) After 5ng/ml TGFβ 1 treatment, the decrease expression of E-Cadherin and the increase expression of vimentin were detected in LNCaP and CWR22rv1 cells in a time dependent manner by Western blot. (b) EMT transition induced by TGFβ 1 could be blocked by CD44 siRNA in LNCaP and CWR22rv1 cells. LNCaP and CWR22rv1 cells were treated with 5ng/ml TGFβ 1 with or without CD44 siRNA for 12hrs and 24hrs. The expressions of CD44, E-Cadherin and Vimentin of different treatments were detected by Western blot assay. (c) Invasion ability of LNCaP and CWR22rv1 cells treated with TGFβ 1 (5ng/ml), CD44 siRNA and both treatments were analyzed in Transwell Chamber assay. Quantitation was shown on the right. Significance was defined as p
    Figure Legend Snippet: TGFβ1 altered EMT with enhanced PCa invasion via modulating CD44 (a) After 5ng/ml TGFβ 1 treatment, the decrease expression of E-Cadherin and the increase expression of vimentin were detected in LNCaP and CWR22rv1 cells in a time dependent manner by Western blot. (b) EMT transition induced by TGFβ 1 could be blocked by CD44 siRNA in LNCaP and CWR22rv1 cells. LNCaP and CWR22rv1 cells were treated with 5ng/ml TGFβ 1 with or without CD44 siRNA for 12hrs and 24hrs. The expressions of CD44, E-Cadherin and Vimentin of different treatments were detected by Western blot assay. (c) Invasion ability of LNCaP and CWR22rv1 cells treated with TGFβ 1 (5ng/ml), CD44 siRNA and both treatments were analyzed in Transwell Chamber assay. Quantitation was shown on the right. Significance was defined as p

    Techniques Used: Expressing, Western Blot, Transwell Chamber Assay, Quantitation Assay

    15) Product Images from "Regulation of connexin 43 by interleukin 1β in adult rat cardiac fibroblasts and effects in an adult rat cardiac myocyte: fibroblast co-culture model"

    Article Title: Regulation of connexin 43 by interleukin 1β in adult rat cardiac fibroblasts and effects in an adult rat cardiac myocyte: fibroblast co-culture model

    Journal: Heliyon

    doi: 10.1016/j.heliyon.2019.e03031

    Effect of IL-1β or TGFβ1 alone or in combination on the mRNA expression of CTGF and αSMA in CFs. Quiesced cells were stimulated with IL-1β, TGFβ1 or IL-1β plus TGFβ1 (all at 10 ng/mL) for 24 h. Cells maintained in 0.5% FBS media were used as a control. mRNA levels of (A) CTGF or (B) αSMA was examined. Data represented as relative quantification (RQ) ± rq max normalised to UBC ( n= 3). * P
    Figure Legend Snippet: Effect of IL-1β or TGFβ1 alone or in combination on the mRNA expression of CTGF and αSMA in CFs. Quiesced cells were stimulated with IL-1β, TGFβ1 or IL-1β plus TGFβ1 (all at 10 ng/mL) for 24 h. Cells maintained in 0.5% FBS media were used as a control. mRNA levels of (A) CTGF or (B) αSMA was examined. Data represented as relative quantification (RQ) ± rq max normalised to UBC ( n= 3). * P

    Techniques Used: Expressing

    Effect of proinflammatory mediators on Cx43 expression in CFs. (A) Quiesced cells were stimulated with 10% FBS, Ang II (500nM), TGFβ1, IL-1β, IL-1α, TNFα (all 10ng/mL) or IL-6 in the presence or absence of the sIL-6R (both 50 ng/mL) for 24 h. Cells maintained in 0.5% FBS media were used as a control. Data represented as relative quantification (RQ) ± rq max normalised to UBC (n = 3). * P
    Figure Legend Snippet: Effect of proinflammatory mediators on Cx43 expression in CFs. (A) Quiesced cells were stimulated with 10% FBS, Ang II (500nM), TGFβ1, IL-1β, IL-1α, TNFα (all 10ng/mL) or IL-6 in the presence or absence of the sIL-6R (both 50 ng/mL) for 24 h. Cells maintained in 0.5% FBS media were used as a control. Data represented as relative quantification (RQ) ± rq max normalised to UBC (n = 3). * P

    Techniques Used: Expressing

    16) Product Images from "SRGN-TGFβ2 regulatory loop confers invasion and metastasis in triple-negative breast cancer"

    Article Title: SRGN-TGFβ2 regulatory loop confers invasion and metastasis in triple-negative breast cancer

    Journal: Oncogenesis

    doi: 10.1038/oncsis.2017.53

    SRGN induces TGFβ2 expression and activates EMT formation. ( a ) Representative western blots of EMT-related markers in MDA-MB-231/shSRGN2#, MDA-MB-231/shNC, MCF7/NC and MCF7/SRGN stable cells. ( b ) Real-time PCR of TGFβ1 and TGFβ2 expression after interference and overexpression of SRGN. * P
    Figure Legend Snippet: SRGN induces TGFβ2 expression and activates EMT formation. ( a ) Representative western blots of EMT-related markers in MDA-MB-231/shSRGN2#, MDA-MB-231/shNC, MCF7/NC and MCF7/SRGN stable cells. ( b ) Real-time PCR of TGFβ1 and TGFβ2 expression after interference and overexpression of SRGN. * P

    Techniques Used: Expressing, Western Blot, Multiple Displacement Amplification, Real-time Polymerase Chain Reaction, Over Expression

    17) Product Images from "Gene expression signature of c-MYC-immortalized human fibroblasts reveals loss of growth inhibitory response to TGF?"

    Article Title: Gene expression signature of c-MYC-immortalized human fibroblasts reveals loss of growth inhibitory response to TGF?

    Journal: Cell Cycle

    doi: 10.4161/cc.10.15.16309

    Disruption of TGFβ signaling in iMYC cells. (A) Empty vector control (pB) cells were plated, treated with 0.1 ng/ml or 0.2 ng/ml TGFβ1 ligand and counted at 2 d and at 4 d. A growth curve showing cell number on the indicated days is shown. (B) eMYC cells were plated and treated as in (A). (C) iMYC cells were plated and treated as in (A).
    Figure Legend Snippet: Disruption of TGFβ signaling in iMYC cells. (A) Empty vector control (pB) cells were plated, treated with 0.1 ng/ml or 0.2 ng/ml TGFβ1 ligand and counted at 2 d and at 4 d. A growth curve showing cell number on the indicated days is shown. (B) eMYC cells were plated and treated as in (A). (C) iMYC cells were plated and treated as in (A).

    Techniques Used: Plasmid Preparation

    p27 protein turnover contributes to the differential growth response of eMYC and iMYC cells to TGFβ. (A) Western analysis of protein lysates from empty vector control (pB), eMYC (MYC) and iMYC (iMYC) cells treated with TGFβ1 ligand. Levels of p27 and GAPDH are shown. (B) Western analysis of protein lysates from empty vector control (pB) and eMYC (MYC) cells transfected with either non-targeting siRNA (−) or against p27 (+). Levels of p27 and GAPDH are shown. (C) eMYC cells transfected with siRNA against p27 were plated 24 h after transfection, treated with 0.1 ng/ml TGFβ1 ligand and counted at 3 d after treatment. Levels of growth in TGFβ1-untreated and treated cells are shown relative to levels of growth in eMYC cells transfected with control siRNA and treated with ligand. (D) Western analysis of protein lysates from empty vector control (pB), eMYC (eMYC) and iMYC (iMYC) cells treated with TGFβ1 ligand, MG132, or both 2 h prior to protein harvest. Levels of p27 and GAPDH are shown.
    Figure Legend Snippet: p27 protein turnover contributes to the differential growth response of eMYC and iMYC cells to TGFβ. (A) Western analysis of protein lysates from empty vector control (pB), eMYC (MYC) and iMYC (iMYC) cells treated with TGFβ1 ligand. Levels of p27 and GAPDH are shown. (B) Western analysis of protein lysates from empty vector control (pB) and eMYC (MYC) cells transfected with either non-targeting siRNA (−) or against p27 (+). Levels of p27 and GAPDH are shown. (C) eMYC cells transfected with siRNA against p27 were plated 24 h after transfection, treated with 0.1 ng/ml TGFβ1 ligand and counted at 3 d after treatment. Levels of growth in TGFβ1-untreated and treated cells are shown relative to levels of growth in eMYC cells transfected with control siRNA and treated with ligand. (D) Western analysis of protein lysates from empty vector control (pB), eMYC (eMYC) and iMYC (iMYC) cells treated with TGFβ1 ligand, MG132, or both 2 h prior to protein harvest. Levels of p27 and GAPDH are shown.

    Techniques Used: Western Blot, Plasmid Preparation, Transfection

    18) Product Images from "OOPHORECTOMY-INDUCED BONE LOSS IS ATTENUATED IN MAGP1-DEFICIENT MICE"

    Article Title: OOPHORECTOMY-INDUCED BONE LOSS IS ATTENUATED IN MAGP1-DEFICIENT MICE

    Journal: Journal of cellular biochemistry

    doi: 10.1002/jcb.23331

    MAGP1 binds TGFβ1 and regulates its bioavailability. 4a) Surface plasmon resonance (BIAcore) was utilized to determine MAGP1’s affinity for pro-osteoclastogenic factors TGFβ1, RANKL, TNFα. 4b-c) RFL-6 cells were stably
    Figure Legend Snippet: MAGP1 binds TGFβ1 and regulates its bioavailability. 4a) Surface plasmon resonance (BIAcore) was utilized to determine MAGP1’s affinity for pro-osteoclastogenic factors TGFβ1, RANKL, TNFα. 4b-c) RFL-6 cells were stably

    Techniques Used: SPR Assay, Stable Transfection

    19) Product Images from "Novel regulatory role of neuropilin-1 in endothelial-to-mesenchymal transition and fibrosis in pancreatic ductal adenocarcinoma"

    Article Title: Novel regulatory role of neuropilin-1 in endothelial-to-mesenchymal transition and fibrosis in pancreatic ductal adenocarcinoma

    Journal: Oncotarget

    doi: 10.18632/oncotarget.11060

    Loss of NRP-1 inhibits TGFβ1- induced EndMT in HUVECs. HUVECs were cultured and transfected with 5 nM of either siNRP-1 or scramble control Total RNA and protein was extracted from the transfected HUVECs at 24, 48 and 72 h post-transfection. ( A ) qPCR data demonstrate successful silencing of NRP-1 (~85% reduction) in siNRP-1 transfected HUVECs after 24 h. All qPCR data are presented as fold change to the scrambled control. ( B ) NRP-1 silencing was confirmed at protein level by immunoblotting at 24, 48 and 72 h after transfection. GAPDH was used as a loading control. ( C ) Representative images from capillary-like tube formation assay demonstrating reduced number of tubes in siNRP-1 compared to scramble control at 6 h. ( D ) Tube formation ability of HUVECs on Matrigel TM was quantified as number of nodes and number of tubes, showing that siNRP-1 caused a significant anti-angiogenic effect as compared to scramble control. * p
    Figure Legend Snippet: Loss of NRP-1 inhibits TGFβ1- induced EndMT in HUVECs. HUVECs were cultured and transfected with 5 nM of either siNRP-1 or scramble control Total RNA and protein was extracted from the transfected HUVECs at 24, 48 and 72 h post-transfection. ( A ) qPCR data demonstrate successful silencing of NRP-1 (~85% reduction) in siNRP-1 transfected HUVECs after 24 h. All qPCR data are presented as fold change to the scrambled control. ( B ) NRP-1 silencing was confirmed at protein level by immunoblotting at 24, 48 and 72 h after transfection. GAPDH was used as a loading control. ( C ) Representative images from capillary-like tube formation assay demonstrating reduced number of tubes in siNRP-1 compared to scramble control at 6 h. ( D ) Tube formation ability of HUVECs on Matrigel TM was quantified as number of nodes and number of tubes, showing that siNRP-1 caused a significant anti-angiogenic effect as compared to scramble control. * p

    Techniques Used: Cell Culture, Transfection, Real-time Polymerase Chain Reaction, Capillary Tube Formation Assay

    Lentivirus-mediated NRP-1 overexpression exacerbates TGFβ1-induced EndMT in HUVECs HUVECs were cultured and transduced with either lentiNRP-1 or blank/empty control lentivirus (lentiControl). Total RNA and protein was extracted from the transduced HUVECs at 24, 48 and 72 h post-transduction. ( A ) qPCR data demonstrate successful overexpression of NRP-1 in lentiNRP-1 transduced HUVECs after 48 h. All qPCR data are presented as fold change to the scrambled control. ( B ) NRP-1 overexpression was confirmed at protein level by immunoblotting. ( C ) Representative images from capillary-like tube formation assay demonstrating increased number of tubes in lentiNRP-1 compared to lentiControl at 6 h time point. ( D ) Tube formation ability of HUVECs on Matrigel TM was quantified as number of nodes and number of tubes and showed that lentiNRP-1 caused a significant pro-angiogenic effect as compared to lentiControl. * p
    Figure Legend Snippet: Lentivirus-mediated NRP-1 overexpression exacerbates TGFβ1-induced EndMT in HUVECs HUVECs were cultured and transduced with either lentiNRP-1 or blank/empty control lentivirus (lentiControl). Total RNA and protein was extracted from the transduced HUVECs at 24, 48 and 72 h post-transduction. ( A ) qPCR data demonstrate successful overexpression of NRP-1 in lentiNRP-1 transduced HUVECs after 48 h. All qPCR data are presented as fold change to the scrambled control. ( B ) NRP-1 overexpression was confirmed at protein level by immunoblotting. ( C ) Representative images from capillary-like tube formation assay demonstrating increased number of tubes in lentiNRP-1 compared to lentiControl at 6 h time point. ( D ) Tube formation ability of HUVECs on Matrigel TM was quantified as number of nodes and number of tubes and showed that lentiNRP-1 caused a significant pro-angiogenic effect as compared to lentiControl. * p

    Techniques Used: Over Expression, Cell Culture, Transduction, Real-time Polymerase Chain Reaction, Capillary Tube Formation Assay

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    Recombinant:

    Article Title: Nintedanib selectively inhibits the activation and tumour-promoting effects of fibroblasts from lung adenocarcinoma patients
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    Article Title: 20S-Hydroxyvitamin D3, Noncalcemic Product of CYP11A1 Action on Vitamin D3, Exhibits Potent Antifibrogenic Activity in Vivo
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    Article Title: Transforming Growth Factor-? (TGF-?1) Activates TAK1 via TAB1-mediated Autophosphorylation, Independent of TGF-? Receptor Kinase Activity in Mesangial Cells *
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    Article Title: CTGF directs fibroblast differentiation from human mesenchymal stem/stromal cells and defines connective tissue healing in a rodent injury model
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    Cell Culture:

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    Concentration Assay:

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  • 95
    R&D Systems tgfβ1 specific inhibitory antibody
    The RGE mutation does not affect Tgfb1 gene expression, LAP inhibitory function, or latent <t>TGFβ1</t> processing and secretion. Serum TGFβ1 levels in Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice before (A) and after (B) removal of a Neo sequence within the mutated Tgfb1 allele. (C) TGFβ levels in medium conditioned by lung fibroblasts derived from Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice. Effects of an anti-TGFβ antibody active against all three TGFβ isoforms (α-TGFβ) and of an antibody that inhibits only TGFβ1 (α-TGFβ1) are shown. Error bars indicate means ± SEM. (D) Tgfb1 gene expression was assessed by semiquantitative RT-PCR using RNA isolated from Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE lung fibroblasts. Reverse-transcribed β-actin mRNA was amplified as a control. (E) Recombinant wild-type and RGE mutant LAP inhibit equally the activity of recombinant TGFβ1, measured with a cell-based luciferase assay. Luciferase activity, expressed as relative light units (RLU), is proportional to TGFβ activity. (F) Wild-type and RGE mutant TGFβ1 cDNAs were expressed in LTBP1S-expressing CHO cells. Secreted proteins were separated by SDS-PAGE in nonreducing conditions and probed with an anti-LAP antibody. Large latent complex (LLC) indicates migration of the LAP–LTBP1 complex.
    Tgfβ1 Specific Inhibitory 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
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    93
    R&D Systems human tgfβ1
    BMP-dependent FGF signaling in lens cells requires active BMP receptors. (A) Dorsomorphin is a specific inhibitor of BMP signaling in DCDMLs. Cultures were treated for 45 min without factors (–) or with 5 ng/ml BMP4, 4 ng/ml <t>TGFβ1,</t> or 10 ng/ml FGF2. Where indicated, cells were pretreated with 5 μM dorsomorphin (DM) before addition of the factor. Whole-cell lysates were probed with antibodies specific for the phosphorylated (activated) forms of Smad1/5, Smad3, or ERK. Representative of three independent experiments; inhibition of either Smad3 or ERK activation never exceeded 20%. (B, C) Dorsomorphin blocks both BMP- and FGF-induced processes in DCDMLs. DCDMLs were incubated without growth factor or with 10 ng/ml BMP4 or FGF2 in either the absence or presence of dorsomorphin as indicated. (B) After 6 d of culture, cells were assayed for synthesis of the fiber differentiation markers δ-crystallin (by [ 35 S]methionine labeling) and CP49 (by quantitative anti-CP49 Western blotting). Fold increase over control ± SD given for each condition ( n = 5). (C) Cultures were assessed for their ability to activate ERK in response to an 8-h incubation with FGF2 as in Figure 1 A. Percentage decrease in activation of ERK in response to FGF in dorsomorphin-treated cells relative to cells not cultured with dorsomorphin was 65 ± 7.7% ( n = 3).
    Human Tgfβ1, 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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    99
    R&D Systems recombinant human tgfβ1
    The Z-cad sensor identifies differential plasticity in HMLER EMT subpopulations upon <t>TGFβ1</t> exposure. a HMLER cells cultured for 24 days with TGFβ1 were sorted into EMT and Bulk –EMT groups and allowed to grow in the absence of TGFβ1 for 13 days. b Parental HMLER cells were grown sorted into EMT and Bulk –EMT groups and allowed to grow for 13 days. c HMLER EMT cells were grown in the presence of the indicated concentrations of decitabine (or vehicle) for 5 days. Medium was changed daily with fresh decitabine. Flow cytometry was performed and the EMT fluorescence signature is indicated ( n = 3 biological replicates for vehicle and 500 nM; n = 2 biological replicates for 100 nM). d qPCR analysis for the indicated genes at different decitabine concentrations is shown. Vehicle treated values were set to 1.0 for each gene. Each concentration was compared to vehicle and analyzed using an unpaired Student’s t test ( n = 3 biological replicates per group). * p value
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    The RGE mutation does not affect Tgfb1 gene expression, LAP inhibitory function, or latent TGFβ1 processing and secretion. Serum TGFβ1 levels in Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice before (A) and after (B) removal of a Neo sequence within the mutated Tgfb1 allele. (C) TGFβ levels in medium conditioned by lung fibroblasts derived from Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice. Effects of an anti-TGFβ antibody active against all three TGFβ isoforms (α-TGFβ) and of an antibody that inhibits only TGFβ1 (α-TGFβ1) are shown. Error bars indicate means ± SEM. (D) Tgfb1 gene expression was assessed by semiquantitative RT-PCR using RNA isolated from Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE lung fibroblasts. Reverse-transcribed β-actin mRNA was amplified as a control. (E) Recombinant wild-type and RGE mutant LAP inhibit equally the activity of recombinant TGFβ1, measured with a cell-based luciferase assay. Luciferase activity, expressed as relative light units (RLU), is proportional to TGFβ activity. (F) Wild-type and RGE mutant TGFβ1 cDNAs were expressed in LTBP1S-expressing CHO cells. Secreted proteins were separated by SDS-PAGE in nonreducing conditions and probed with an anti-LAP antibody. Large latent complex (LLC) indicates migration of the LAP–LTBP1 complex.

    Journal: The Journal of Cell Biology

    Article Title: Absence of integrin-mediated TGF?1 activation in vivo recapitulates the phenotype of TGF?1-null mice

    doi: 10.1083/jcb.200611044

    Figure Lengend Snippet: The RGE mutation does not affect Tgfb1 gene expression, LAP inhibitory function, or latent TGFβ1 processing and secretion. Serum TGFβ1 levels in Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice before (A) and after (B) removal of a Neo sequence within the mutated Tgfb1 allele. (C) TGFβ levels in medium conditioned by lung fibroblasts derived from Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice. Effects of an anti-TGFβ antibody active against all three TGFβ isoforms (α-TGFβ) and of an antibody that inhibits only TGFβ1 (α-TGFβ1) are shown. Error bars indicate means ± SEM. (D) Tgfb1 gene expression was assessed by semiquantitative RT-PCR using RNA isolated from Tgfb1 −/− , Tgfb1 +/RGE , and Tgfb1 RGE/RGE lung fibroblasts. Reverse-transcribed β-actin mRNA was amplified as a control. (E) Recombinant wild-type and RGE mutant LAP inhibit equally the activity of recombinant TGFβ1, measured with a cell-based luciferase assay. Luciferase activity, expressed as relative light units (RLU), is proportional to TGFβ activity. (F) Wild-type and RGE mutant TGFβ1 cDNAs were expressed in LTBP1S-expressing CHO cells. Secreted proteins were separated by SDS-PAGE in nonreducing conditions and probed with an anti-LAP antibody. Large latent complex (LLC) indicates migration of the LAP–LTBP1 complex.

    Article Snippet: Samples were tested in the presence and absence of a TGFβ1-specific inhibitory antibody (R & D Systems) or an inhibitory antibody against all three TGFβ isoforms (1D11; R & D Systems).

    Techniques: Mutagenesis, Expressing, Mouse Assay, Sequencing, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Isolation, Amplification, Recombinant, Activity Assay, Luciferase, SDS Page, Migration

    Generation of mice with a targeted mutation of Tgfb1 . (A) Schematic of Tgfb1 (top) and fragments thereof used for insertion in targeting vector (bottom). Box shows mutations (solid underlines) introduced in exon 5 to encode a BstUI restriction site (dashed underline) and the D-to-E mutation. Restriction sites: A, ApaI; B, BamHI; H, HindIII; S, SalI. Neo , neomycin resistance cassette; TK , thymidine kinase cassette. (B) The TGFβ1 mRNA encodes a protein sequence consisting of a signal peptide (SP), propeptide (LAP), and the TGFβ1 cytokine (top). The integrin-binding motif RGD is near the C terminus of LAP. The processed latent factor consists of noncovalently associated disulfide-linked homodimers of the LAP and TGFβ1 monomers (bottom). LAP can be disulfide linked via cys-33 to one of the cysteine-repeat domains (ovals) of LTBP-1, -3, or -4. (C) PCR genotyping results from Tgfb1 +/+ , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice.

    Journal: The Journal of Cell Biology

    Article Title: Absence of integrin-mediated TGF?1 activation in vivo recapitulates the phenotype of TGF?1-null mice

    doi: 10.1083/jcb.200611044

    Figure Lengend Snippet: Generation of mice with a targeted mutation of Tgfb1 . (A) Schematic of Tgfb1 (top) and fragments thereof used for insertion in targeting vector (bottom). Box shows mutations (solid underlines) introduced in exon 5 to encode a BstUI restriction site (dashed underline) and the D-to-E mutation. Restriction sites: A, ApaI; B, BamHI; H, HindIII; S, SalI. Neo , neomycin resistance cassette; TK , thymidine kinase cassette. (B) The TGFβ1 mRNA encodes a protein sequence consisting of a signal peptide (SP), propeptide (LAP), and the TGFβ1 cytokine (top). The integrin-binding motif RGD is near the C terminus of LAP. The processed latent factor consists of noncovalently associated disulfide-linked homodimers of the LAP and TGFβ1 monomers (bottom). LAP can be disulfide linked via cys-33 to one of the cysteine-repeat domains (ovals) of LTBP-1, -3, or -4. (C) PCR genotyping results from Tgfb1 +/+ , Tgfb1 +/RGE , and Tgfb1 RGE/RGE mice.

    Article Snippet: Samples were tested in the presence and absence of a TGFβ1-specific inhibitory antibody (R & D Systems) or an inhibitory antibody against all three TGFβ isoforms (1D11; R & D Systems).

    Techniques: Mouse Assay, Mutagenesis, Plasmid Preparation, Sequencing, Binding Assay, Polymerase Chain Reaction

    BMP-dependent FGF signaling in lens cells requires active BMP receptors. (A) Dorsomorphin is a specific inhibitor of BMP signaling in DCDMLs. Cultures were treated for 45 min without factors (–) or with 5 ng/ml BMP4, 4 ng/ml TGFβ1, or 10 ng/ml FGF2. Where indicated, cells were pretreated with 5 μM dorsomorphin (DM) before addition of the factor. Whole-cell lysates were probed with antibodies specific for the phosphorylated (activated) forms of Smad1/5, Smad3, or ERK. Representative of three independent experiments; inhibition of either Smad3 or ERK activation never exceeded 20%. (B, C) Dorsomorphin blocks both BMP- and FGF-induced processes in DCDMLs. DCDMLs were incubated without growth factor or with 10 ng/ml BMP4 or FGF2 in either the absence or presence of dorsomorphin as indicated. (B) After 6 d of culture, cells were assayed for synthesis of the fiber differentiation markers δ-crystallin (by [ 35 S]methionine labeling) and CP49 (by quantitative anti-CP49 Western blotting). Fold increase over control ± SD given for each condition ( n = 5). (C) Cultures were assessed for their ability to activate ERK in response to an 8-h incubation with FGF2 as in Figure 1 A. Percentage decrease in activation of ERK in response to FGF in dorsomorphin-treated cells relative to cells not cultured with dorsomorphin was 65 ± 7.7% ( n = 3).

    Journal: Molecular Biology of the Cell

    Article Title: Synergistic interaction between the fibroblast growth factor and bone morphogenetic protein signaling pathways in lens cells

    doi: 10.1091/mbc.E15-02-0117

    Figure Lengend Snippet: BMP-dependent FGF signaling in lens cells requires active BMP receptors. (A) Dorsomorphin is a specific inhibitor of BMP signaling in DCDMLs. Cultures were treated for 45 min without factors (–) or with 5 ng/ml BMP4, 4 ng/ml TGFβ1, or 10 ng/ml FGF2. Where indicated, cells were pretreated with 5 μM dorsomorphin (DM) before addition of the factor. Whole-cell lysates were probed with antibodies specific for the phosphorylated (activated) forms of Smad1/5, Smad3, or ERK. Representative of three independent experiments; inhibition of either Smad3 or ERK activation never exceeded 20%. (B, C) Dorsomorphin blocks both BMP- and FGF-induced processes in DCDMLs. DCDMLs were incubated without growth factor or with 10 ng/ml BMP4 or FGF2 in either the absence or presence of dorsomorphin as indicated. (B) After 6 d of culture, cells were assayed for synthesis of the fiber differentiation markers δ-crystallin (by [ 35 S]methionine labeling) and CP49 (by quantitative anti-CP49 Western blotting). Fold increase over control ± SD given for each condition ( n = 5). (C) Cultures were assessed for their ability to activate ERK in response to an 8-h incubation with FGF2 as in Figure 1 A. Percentage decrease in activation of ERK in response to FGF in dorsomorphin-treated cells relative to cells not cultured with dorsomorphin was 65 ± 7.7% ( n = 3).

    Article Snippet: Materials Recombinant bovine FGF2, human FGF9, human TGFβ1, mouse noggin/Fc chimera, human BMP4, human BMP6, and anti-BMP antibody (MAB3552) were from R & D Systems (Minneapolis, MN).

    Techniques: Inhibition, Activation Assay, Incubation, Labeling, Western Blot, Cell Culture

    FGF specifically increases expression of the BMP signaling reporter BRE-Luc in lens cells. (A–E) DCDMLs were transfected with plasmids encoding BRE-Luc (A–C), SBE4-Luc (D), or 5′ Del 6 Id1-Luc (E) and cultured with no additions (control), 5 ng/ml BMP4, 10 ng/ml FGF2, 1 ng/ml FGF2, 4 ng/ml TGFβ1, or 50 nM TPA for 22 h (A, B, D, E), or 2 h (C). Where indicated, the FGFR inhibitor PD173074 (PD) was also present. Cultures were either fixed and immunostained with anti-luciferase antibodies (A, D, E) or lysed and equal amount of total cell protein analyzed by Western blot using the same antibody (B, C). Representative of three or more independent experiments. (F) Experiments in which a plasmid encoding constitutively expressed EGFP was cotransfected with 5′ Del 6 Id1-Luc at a ratio of 1:4 demonstrated equal levels of expression of EGFP when normalized to either total protein or tubulin, indicating that the lack of expression of 5′ Del 6 Id1-Luc in E was not due to a lack of cell transfection.

    Journal: Molecular Biology of the Cell

    Article Title: Synergistic interaction between the fibroblast growth factor and bone morphogenetic protein signaling pathways in lens cells

    doi: 10.1091/mbc.E15-02-0117

    Figure Lengend Snippet: FGF specifically increases expression of the BMP signaling reporter BRE-Luc in lens cells. (A–E) DCDMLs were transfected with plasmids encoding BRE-Luc (A–C), SBE4-Luc (D), or 5′ Del 6 Id1-Luc (E) and cultured with no additions (control), 5 ng/ml BMP4, 10 ng/ml FGF2, 1 ng/ml FGF2, 4 ng/ml TGFβ1, or 50 nM TPA for 22 h (A, B, D, E), or 2 h (C). Where indicated, the FGFR inhibitor PD173074 (PD) was also present. Cultures were either fixed and immunostained with anti-luciferase antibodies (A, D, E) or lysed and equal amount of total cell protein analyzed by Western blot using the same antibody (B, C). Representative of three or more independent experiments. (F) Experiments in which a plasmid encoding constitutively expressed EGFP was cotransfected with 5′ Del 6 Id1-Luc at a ratio of 1:4 demonstrated equal levels of expression of EGFP when normalized to either total protein or tubulin, indicating that the lack of expression of 5′ Del 6 Id1-Luc in E was not due to a lack of cell transfection.

    Article Snippet: Materials Recombinant bovine FGF2, human FGF9, human TGFβ1, mouse noggin/Fc chimera, human BMP4, human BMP6, and anti-BMP antibody (MAB3552) were from R & D Systems (Minneapolis, MN).

    Techniques: Expressing, Transfection, Cell Culture, Luciferase, Western Blot, Plasmid Preparation

    The Z-cad sensor identifies differential plasticity in HMLER EMT subpopulations upon TGFβ1 exposure. a HMLER cells cultured for 24 days with TGFβ1 were sorted into EMT and Bulk –EMT groups and allowed to grow in the absence of TGFβ1 for 13 days. b Parental HMLER cells were grown sorted into EMT and Bulk –EMT groups and allowed to grow for 13 days. c HMLER EMT cells were grown in the presence of the indicated concentrations of decitabine (or vehicle) for 5 days. Medium was changed daily with fresh decitabine. Flow cytometry was performed and the EMT fluorescence signature is indicated ( n = 3 biological replicates for vehicle and 500 nM; n = 2 biological replicates for 100 nM). d qPCR analysis for the indicated genes at different decitabine concentrations is shown. Vehicle treated values were set to 1.0 for each gene. Each concentration was compared to vehicle and analyzed using an unpaired Student’s t test ( n = 3 biological replicates per group). * p value

    Journal: BMC Biology

    Article Title: The Z-cad dual fluorescent sensor detects dynamic changes between the epithelial and mesenchymal cellular states

    doi: 10.1186/s12915-016-0269-y

    Figure Lengend Snippet: The Z-cad sensor identifies differential plasticity in HMLER EMT subpopulations upon TGFβ1 exposure. a HMLER cells cultured for 24 days with TGFβ1 were sorted into EMT and Bulk –EMT groups and allowed to grow in the absence of TGFβ1 for 13 days. b Parental HMLER cells were grown sorted into EMT and Bulk –EMT groups and allowed to grow for 13 days. c HMLER EMT cells were grown in the presence of the indicated concentrations of decitabine (or vehicle) for 5 days. Medium was changed daily with fresh decitabine. Flow cytometry was performed and the EMT fluorescence signature is indicated ( n = 3 biological replicates for vehicle and 500 nM; n = 2 biological replicates for 100 nM). d qPCR analysis for the indicated genes at different decitabine concentrations is shown. Vehicle treated values were set to 1.0 for each gene. Each concentration was compared to vehicle and analyzed using an unpaired Student’s t test ( n = 3 biological replicates per group). * p value

    Article Snippet: For EMT induction HMLER cells were treated with 5 ng/mL recombinant human TGFβ1 (R & D Systems, 240-B).

    Techniques: Cell Culture, Flow Cytometry, Cytometry, Fluorescence, Real-time Polymerase Chain Reaction, Concentration Assay

    The Z-cad sensor identifies EMT induced by TGFβ1 in HMLER cells. a F-actin stress fibers increase during TGFβ1 compared to vehicle treatment (26 days) indicating EMT. Scale bar = 100 μm. b RNA analysis by qRT-PCR at indicated time points during vehicle or TGFβ1 treatment ( n = 3 biological replicates per time point). Vehicle-treated values were set to 1.0 for each gene and analyzed using an unpaired Student’s t test. c Fluorescent confocal microscopy indicating a gain of GFP and loss of RFP at 14 days of TGFβ1 versus vehicle treatment. Mesenchymal-appearing bright GFP cells are indicated by arrows . Scale bar = 20 μm. d Flow cytometric analysis of HMLER cells containing Z-cad sensor showing an increase in cells displaying GFP hi RFP low/neg EMT fluorescence signature during TGFβ1 treatment at the times indicated. e Quantitation of cells displaying the EMT signature in HMLER cells containing Z-cad dual sensor from d . Bar graphs depict the percentage of cells displaying the EMT fluorescence signature in TGFβ1- versus vehicle-treated groups ( n = 3 biological replicates per time point). Data were analyzed using an unpaired Student’s t test. * p value

    Journal: BMC Biology

    Article Title: The Z-cad dual fluorescent sensor detects dynamic changes between the epithelial and mesenchymal cellular states

    doi: 10.1186/s12915-016-0269-y

    Figure Lengend Snippet: The Z-cad sensor identifies EMT induced by TGFβ1 in HMLER cells. a F-actin stress fibers increase during TGFβ1 compared to vehicle treatment (26 days) indicating EMT. Scale bar = 100 μm. b RNA analysis by qRT-PCR at indicated time points during vehicle or TGFβ1 treatment ( n = 3 biological replicates per time point). Vehicle-treated values were set to 1.0 for each gene and analyzed using an unpaired Student’s t test. c Fluorescent confocal microscopy indicating a gain of GFP and loss of RFP at 14 days of TGFβ1 versus vehicle treatment. Mesenchymal-appearing bright GFP cells are indicated by arrows . Scale bar = 20 μm. d Flow cytometric analysis of HMLER cells containing Z-cad sensor showing an increase in cells displaying GFP hi RFP low/neg EMT fluorescence signature during TGFβ1 treatment at the times indicated. e Quantitation of cells displaying the EMT signature in HMLER cells containing Z-cad dual sensor from d . Bar graphs depict the percentage of cells displaying the EMT fluorescence signature in TGFβ1- versus vehicle-treated groups ( n = 3 biological replicates per time point). Data were analyzed using an unpaired Student’s t test. * p value

    Article Snippet: For EMT induction HMLER cells were treated with 5 ng/mL recombinant human TGFβ1 (R & D Systems, 240-B).

    Techniques: Quantitative RT-PCR, Confocal Microscopy, Flow Cytometry, Fluorescence, Quantitation Assay

    TFAP2A overexpression in NMuMG modulates epithelial plasticity. a Expression of either GFP or TFAP2A was induced by 72 h doxycycline treatment in NMuMG cells stably transduced with either pCLX-GFP or pCLX-TFAP2A. Morphological changes and sparse cell arrangement are visible in phase contrast microscopy upon TFAP2A expression. Scale bar: 50 μm. b Gene expression log 2 fold changes of EMT markers (TFs) were calculated from mRNA-seq samples of doxycycline-induced, TGFβ1-treated (72 h, 2 ng/mL) pCLX-GFP (pCLX-GFP + TGF-beta), doxycycline-induced pCLX-TFAP2A (pCLX-TFAP2A), as well as of doxycycline-induced, TGFβ1-treated (72 h, 2 ng/mL) pCLX-TFAP2A (pCLX-TFAP2A + TGF-beta) cell lines relative to doxycycline-induced pCLX-GFP (pCLX-GFP) cell line. Shown are the mean log 2 fold changes (+/- 1 standard deviation) from two experiments. TFAP2A overexpression is apparent in both TFAP2A-induced samples ( dark green and dark blue ) but is not induced in cells treated with TGFβ1 alone ( light blue ). The EMT-inducing TFs have increased expression upon TFAP2A induction. * indicates a p -value ≤ 0.05 and ** a p -value ≤ 0.01 in a two-tailed t -test. c The transcriptional activities of TFAP2{A,C} and SNAI1..3 motifs in different conditions, as inferred with ISMARA from mRNA-seq data as described in ( b ). The two replicates from each condition are plotted next to each other

    Journal: Biology Direct

    Article Title: TFAP2A is a component of the ZEB1/2 network that regulates TGFB1-induced epithelial to mesenchymal transition

    doi: 10.1186/s13062-017-0180-7

    Figure Lengend Snippet: TFAP2A overexpression in NMuMG modulates epithelial plasticity. a Expression of either GFP or TFAP2A was induced by 72 h doxycycline treatment in NMuMG cells stably transduced with either pCLX-GFP or pCLX-TFAP2A. Morphological changes and sparse cell arrangement are visible in phase contrast microscopy upon TFAP2A expression. Scale bar: 50 μm. b Gene expression log 2 fold changes of EMT markers (TFs) were calculated from mRNA-seq samples of doxycycline-induced, TGFβ1-treated (72 h, 2 ng/mL) pCLX-GFP (pCLX-GFP + TGF-beta), doxycycline-induced pCLX-TFAP2A (pCLX-TFAP2A), as well as of doxycycline-induced, TGFβ1-treated (72 h, 2 ng/mL) pCLX-TFAP2A (pCLX-TFAP2A + TGF-beta) cell lines relative to doxycycline-induced pCLX-GFP (pCLX-GFP) cell line. Shown are the mean log 2 fold changes (+/- 1 standard deviation) from two experiments. TFAP2A overexpression is apparent in both TFAP2A-induced samples ( dark green and dark blue ) but is not induced in cells treated with TGFβ1 alone ( light blue ). The EMT-inducing TFs have increased expression upon TFAP2A induction. * indicates a p -value ≤ 0.05 and ** a p -value ≤ 0.01 in a two-tailed t -test. c The transcriptional activities of TFAP2{A,C} and SNAI1..3 motifs in different conditions, as inferred with ISMARA from mRNA-seq data as described in ( b ). The two replicates from each condition are plotted next to each other

    Article Snippet: Recombinant human TGFβ1 was obtained from R & D Systems.

    Techniques: Over Expression, Expressing, Stable Transfection, Transduction, Microscopy, Standard Deviation, Two Tailed Test

    TFAP2A expression and activity profile in the NMuMG EMT model. a - b NMuMG cells were treated with 2 ng/mL of TGFβ1 for 72 h and were stained for TFAP2A and F-Actin ( a ) and TFAP2A and E-cadherin ( b ). The merged panels represent colocalization of the imaged markers with the nucleus which was stained with DAPI and compared to controls. Scale bar represents 50 μm. c NMuMG cells were treated for 14 days with 2 ng/mL of TGFβ1. Quantitative RT-PCR of Tfap2a during the time course of this treatment indicates that Tfap2a mRNA levels are reduced upon EMT. The EMT markers E-cadherin (Cdh1), Fibronectin (Fn1), Occludin (Ocln), and Vimentin (Vim) follow the expected trend. d Two mRNA-seq samples from independent wells were prepared from a time course of NMuMG cells treated for 14 days with 2 ng/mL of TGFβ1, and the data was consequently analyzed with ISMARA [ 30 ]. The figure depicts the dynamics of TFAP2A/C transcriptional activity during the time course. The sequence logo of the TFAP2A/C binding motif is also indicated. e - f Lysates from NMuMG/E9 cells treated with 2 ng/mL of TGFβ1 for 72 h were probed for TFAP2A, GAPDH and Lamin B expression by WB and their levels compared with the expression levels of Actin and also to the Ponceau-stained membrane ( e ). The bar plot represents the densitometric quantification of the TFAP2A protein levels upon treatment compared to the control ( f ) ** indicates a p -value

    Journal: Biology Direct

    Article Title: TFAP2A is a component of the ZEB1/2 network that regulates TGFB1-induced epithelial to mesenchymal transition

    doi: 10.1186/s13062-017-0180-7

    Figure Lengend Snippet: TFAP2A expression and activity profile in the NMuMG EMT model. a - b NMuMG cells were treated with 2 ng/mL of TGFβ1 for 72 h and were stained for TFAP2A and F-Actin ( a ) and TFAP2A and E-cadherin ( b ). The merged panels represent colocalization of the imaged markers with the nucleus which was stained with DAPI and compared to controls. Scale bar represents 50 μm. c NMuMG cells were treated for 14 days with 2 ng/mL of TGFβ1. Quantitative RT-PCR of Tfap2a during the time course of this treatment indicates that Tfap2a mRNA levels are reduced upon EMT. The EMT markers E-cadherin (Cdh1), Fibronectin (Fn1), Occludin (Ocln), and Vimentin (Vim) follow the expected trend. d Two mRNA-seq samples from independent wells were prepared from a time course of NMuMG cells treated for 14 days with 2 ng/mL of TGFβ1, and the data was consequently analyzed with ISMARA [ 30 ]. The figure depicts the dynamics of TFAP2A/C transcriptional activity during the time course. The sequence logo of the TFAP2A/C binding motif is also indicated. e - f Lysates from NMuMG/E9 cells treated with 2 ng/mL of TGFβ1 for 72 h were probed for TFAP2A, GAPDH and Lamin B expression by WB and their levels compared with the expression levels of Actin and also to the Ponceau-stained membrane ( e ). The bar plot represents the densitometric quantification of the TFAP2A protein levels upon treatment compared to the control ( f ) ** indicates a p -value

    Article Snippet: Recombinant human TGFβ1 was obtained from R & D Systems.

    Techniques: Expressing, Activity Assay, Staining, Quantitative RT-PCR, Sequencing, Binding Assay, Western Blot