epidermal growth factor egf  (PeproTech)

 
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    Name:
    Recombinant Murine EGF
    Description:
    Epidermal Growth Factor EGF is a potent growth factor that stimulates the proliferation of various epidermal and epithelial cells Additionally EGF has been shown to inhibit gastric secretion and to be involved in wound healing EGF signals through a receptor known as c erbB which is a class I tyrosine kinase receptor This receptor also binds with TGF α and VGF vaccinia virus growth factor Recombinant murine EGF is a 6 0 kDa globular protein containing 53 amino acid residues including 3 intramolecular disulfide bonds
    Catalog Number:
    315-09-100UG
    Price:
    80.00
    Category:
    Recombinant Proteins
    Source:
    E.coli
    Reactivity:
    Human
    Purity:
    98.0
    Quantity:
    100UG
    		Buy from Supplier

    Structured Review

    PeproTech epidermal growth factor egf
    Vitreous-driven activation of PDGFRα was selective, prolonged, and dependent on ROS. (A) Serum-starved ARPE19α cells were pretreated with NAC (10 mM) for 30 min and then stimulated with buffer (−), with <t>PDGF-A,</t> <t>EGF,</t> or IGF-1 (IGF) (each at 50 ng/ml), or with RV for 10 min. The lanes designated C1 and C2 were stimulated with buffer and RV, respectively. The cells were lysed, and the resulting lysates were immunoprecipitated using antibodies against PDGFRα, EGFR, IGF-1R, or nonimmune IgG (C1 and C2). The resulting immunoprecipitates were subjected to Western blot analysis using an antiphosphotyrosine antibody (pY20). The membranes were reprobed with antibodies against PDGFRα, EGFR, or IGF-1R. The fold values are the p-PDGFRα/PDGFRα ratio, the p-EGFR/EGFR ratio, or the p-IGF-1R/IGF-1R ratio. The data are representative of three experiments; similar results were obtained when the experiment was repeated with primary human corneal fibroblasts instead of ARPE19α cells (data not shown). (B) Serum-deprived ARPE19α cells were pretreated with NAC (10 mM) for 30 min before stimulation with PDGF-A (50 ng/ml) or RV for 10 min. For the 16-h stimulation with PDGF-A or RV, the 30-min NAC (10 mM) treatment was from 15.5 to 16 h. Following stimulation, the cells were lysed and subjected to Western blotting using the indicated antibodies. The fold values are the p-PDGFRα/PDGFRα ratio or the p-Akt/Akt ratio. The blots presented are representative of three experiments. (C) Serum-starved ARPE19α cells were treated with PDGF-A (50 ng/ml) or RV for 10 min, acid washed (to remove the growth factors), and harvested either at 10 min or at 15 h and 50 min later. The lysates were subjected to Western blot analysis using a phospho-PDGFRα Y754 antibody. The membrane was reprobed with a PDGFRα antibody (27P). The fold values are the p-PDGFRα/PDGFRα ratio. The data presented are representative of three experiments.
    Epidermal Growth Factor EGF is a potent growth factor that stimulates the proliferation of various epidermal and epithelial cells Additionally EGF has been shown to inhibit gastric secretion and to be involved in wound healing EGF signals through a receptor known as c erbB which is a class I tyrosine kinase receptor This receptor also binds with TGF α and VGF vaccinia virus growth factor Recombinant murine EGF is a 6 0 kDa globular protein containing 53 amino acid residues including 3 intramolecular disulfide bonds
    https://www.bioz.com/result/epidermal growth factor egf/product/PeproTech
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    epidermal growth factor egf - by Bioz Stars, 2021-04
    97/100 stars

    Images

    1) Product Images from "A Reactive Oxygen Species-Mediated, Self-Perpetuating Loop Persistently Activates Platelet-Derived Growth Factor Receptor ?"

    Article Title: A Reactive Oxygen Species-Mediated, Self-Perpetuating Loop Persistently Activates Platelet-Derived Growth Factor Receptor ?

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.00839-13

    Vitreous-driven activation of PDGFRα was selective, prolonged, and dependent on ROS. (A) Serum-starved ARPE19α cells were pretreated with NAC (10 mM) for 30 min and then stimulated with buffer (−), with PDGF-A, EGF, or IGF-1 (IGF) (each at 50 ng/ml), or with RV for 10 min. The lanes designated C1 and C2 were stimulated with buffer and RV, respectively. The cells were lysed, and the resulting lysates were immunoprecipitated using antibodies against PDGFRα, EGFR, IGF-1R, or nonimmune IgG (C1 and C2). The resulting immunoprecipitates were subjected to Western blot analysis using an antiphosphotyrosine antibody (pY20). The membranes were reprobed with antibodies against PDGFRα, EGFR, or IGF-1R. The fold values are the p-PDGFRα/PDGFRα ratio, the p-EGFR/EGFR ratio, or the p-IGF-1R/IGF-1R ratio. The data are representative of three experiments; similar results were obtained when the experiment was repeated with primary human corneal fibroblasts instead of ARPE19α cells (data not shown). (B) Serum-deprived ARPE19α cells were pretreated with NAC (10 mM) for 30 min before stimulation with PDGF-A (50 ng/ml) or RV for 10 min. For the 16-h stimulation with PDGF-A or RV, the 30-min NAC (10 mM) treatment was from 15.5 to 16 h. Following stimulation, the cells were lysed and subjected to Western blotting using the indicated antibodies. The fold values are the p-PDGFRα/PDGFRα ratio or the p-Akt/Akt ratio. The blots presented are representative of three experiments. (C) Serum-starved ARPE19α cells were treated with PDGF-A (50 ng/ml) or RV for 10 min, acid washed (to remove the growth factors), and harvested either at 10 min or at 15 h and 50 min later. The lysates were subjected to Western blot analysis using a phospho-PDGFRα Y754 antibody. The membrane was reprobed with a PDGFRα antibody (27P). The fold values are the p-PDGFRα/PDGFRα ratio. The data presented are representative of three experiments.
    Figure Legend Snippet: Vitreous-driven activation of PDGFRα was selective, prolonged, and dependent on ROS. (A) Serum-starved ARPE19α cells were pretreated with NAC (10 mM) for 30 min and then stimulated with buffer (−), with PDGF-A, EGF, or IGF-1 (IGF) (each at 50 ng/ml), or with RV for 10 min. The lanes designated C1 and C2 were stimulated with buffer and RV, respectively. The cells were lysed, and the resulting lysates were immunoprecipitated using antibodies against PDGFRα, EGFR, IGF-1R, or nonimmune IgG (C1 and C2). The resulting immunoprecipitates were subjected to Western blot analysis using an antiphosphotyrosine antibody (pY20). The membranes were reprobed with antibodies against PDGFRα, EGFR, or IGF-1R. The fold values are the p-PDGFRα/PDGFRα ratio, the p-EGFR/EGFR ratio, or the p-IGF-1R/IGF-1R ratio. The data are representative of three experiments; similar results were obtained when the experiment was repeated with primary human corneal fibroblasts instead of ARPE19α cells (data not shown). (B) Serum-deprived ARPE19α cells were pretreated with NAC (10 mM) for 30 min before stimulation with PDGF-A (50 ng/ml) or RV for 10 min. For the 16-h stimulation with PDGF-A or RV, the 30-min NAC (10 mM) treatment was from 15.5 to 16 h. Following stimulation, the cells were lysed and subjected to Western blotting using the indicated antibodies. The fold values are the p-PDGFRα/PDGFRα ratio or the p-Akt/Akt ratio. The blots presented are representative of three experiments. (C) Serum-starved ARPE19α cells were treated with PDGF-A (50 ng/ml) or RV for 10 min, acid washed (to remove the growth factors), and harvested either at 10 min or at 15 h and 50 min later. The lysates were subjected to Western blot analysis using a phospho-PDGFRα Y754 antibody. The membrane was reprobed with a PDGFRα antibody (27P). The fold values are the p-PDGFRα/PDGFRα ratio. The data presented are representative of three experiments.

    Techniques Used: Activation Assay, Immunoprecipitation, Western Blot

    2) Product Images from "The Myc 3′ Wnt-Responsive Element Suppresses Colonic Tumorigenesis"

    Article Title: The Myc 3′ Wnt-Responsive Element Suppresses Colonic Tumorigenesis

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.00969-13

    The Myc 3′ WRE is required for mitogen-induced Myc gene expression. The proliferative regions of colonic crypts were isolated from 7-week-old Myc 3′ WRE −/− mice and WT littermates and were not treated (Ctrl.) or treated for 1 or 3 h with recombinant Wnt3A, R-spondin1, and EGF (A), Wnt3A and R-spondin1 (B), or EGF (C). RNAs were isolated, cDNAs were synthesized, and Myc expression levels were evaluated by using quantitative real-time PCR. Myc mRNA levels were normalized to β-actin gene levels ( n = 3 mice analyzed per genotype, with 12 total PCR replicates per sample). Errors are standard errors of the means (***, P
    Figure Legend Snippet: The Myc 3′ WRE is required for mitogen-induced Myc gene expression. The proliferative regions of colonic crypts were isolated from 7-week-old Myc 3′ WRE −/− mice and WT littermates and were not treated (Ctrl.) or treated for 1 or 3 h with recombinant Wnt3A, R-spondin1, and EGF (A), Wnt3A and R-spondin1 (B), or EGF (C). RNAs were isolated, cDNAs were synthesized, and Myc expression levels were evaluated by using quantitative real-time PCR. Myc mRNA levels were normalized to β-actin gene levels ( n = 3 mice analyzed per genotype, with 12 total PCR replicates per sample). Errors are standard errors of the means (***, P

    Techniques Used: Expressing, Isolation, Mouse Assay, Recombinant, Synthesized, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    Mitogens induce an exchange of transcriptional regulatory complexes at Myc 5′ and 3′ Wnt-responsive elements. (A) Diagram of the Myc locus, with the Myc 5′ and 3′ WREs indicated by white rectangles. Gray rectangles indicate the positions of the amplicons produced in quantitative real-time PCRs in panels B to G. (B) The proliferative region of colonic crypts were isolated from 7-week-old WT mice and were untreated (Ctrl.) or treated with recombinant Wnt3A, EGF, and R-spondin1 (mitogens) for 3 h. ChIP assays were performed by using antibodies directed against RNA polymerase 2 (RNAP), and the precipitated DNA was measured by using quantitative real-time PCR with oligonucleotides that hybridized to the regions indicated. Colonic crypts were isolated from 3 mice, and 12 PCR replicates were prepared for each sample. Errors are standard errors of the means (**, P
    Figure Legend Snippet: Mitogens induce an exchange of transcriptional regulatory complexes at Myc 5′ and 3′ Wnt-responsive elements. (A) Diagram of the Myc locus, with the Myc 5′ and 3′ WREs indicated by white rectangles. Gray rectangles indicate the positions of the amplicons produced in quantitative real-time PCRs in panels B to G. (B) The proliferative region of colonic crypts were isolated from 7-week-old WT mice and were untreated (Ctrl.) or treated with recombinant Wnt3A, EGF, and R-spondin1 (mitogens) for 3 h. ChIP assays were performed by using antibodies directed against RNA polymerase 2 (RNAP), and the precipitated DNA was measured by using quantitative real-time PCR with oligonucleotides that hybridized to the regions indicated. Colonic crypts were isolated from 3 mice, and 12 PCR replicates were prepared for each sample. Errors are standard errors of the means (**, P

    Techniques Used: Produced, Isolation, Mouse Assay, Recombinant, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    Related Articles

    Derivative Assay:

    Article Title: Role of Palladin Phosphorylation by Extracellular Signal-Regulated Kinase in Cell Migration
    Article Snippet: Anti-HA antibody was obtained from Roche (Basel, Switzerland), anti-ERK, anti-phospho-tyrosine (PY20) and anti-Myc (9E10) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA), anti-GFP antibody was from Nacalai Tesque (Tokyo, Japan), anti-MPM2 antibody was from Millipore (Billerica, MA), anti-Rac, anti-Cdc42 and anti-Abl antibodies were from BD Biosciences (San Jose, CA), and anti-phospho-ERK antibody was from Cell Signaling (Tokyo, Japan). .. Epidermal Growth Factor (EGF) and Platelet Derived Growth Factor (PDGF) were purchased from PeproTech (Rocky Hill, NJ). .. U0126 and LY294002 were obtained from Wako (Osaka, Japan), and STI571 was from Cayman chemical (Ann Arbor, MI).

    Recombinant:

    Article Title: Cancer stem-like cells persist in established cell lines through autocrine activation of EGFR signaling
    Article Snippet: .. Recombinant EGF and bFGF were obtained from Peprotech (Rocky Hill, NJ, USA). .. Human breast cancer cell line (MCF7), glioma cell line (U87) and non-small cell lung cancer cell line (A549) were cultured in Dulbecco's modified Eagle's medium (DMEM) and RPMI-1640, respectively, supplemented with 10% fetal bovine serum (FBS; JR Scientific, Inc., Woodland, CA, USA) and 1% penicillin/streptomycin and were maintained at 37˚C in a 5% CO2 incubator.

    Article Title: VEGF Stimulates RCAN1.4 Expression in Endothelial Cells via a Pathway Requiring Ca2+/Calcineurin and Protein Kinase C-?
    Article Snippet: Furthermore, RCAN1.4 was required for migration and efficient tubular morphogenesis in these cells. .. Materials Recombinant human VEGF-A165 , epidermal growth factor (EGF), hepatocyte growth factor (HGF) and colony stimulating factor-1 (CSF-1) were obtained from Peprotech. .. Recombinant human VEGF-B167 and recombinant Orf virus VEGF-E were obtained from Promokine.

    Article Title: Inhibition of Epidermal Growth Factor Receptor Tyrosine Kinase Ameliorates Collagen-Induced Arthritis
    Article Snippet: .. For all studies, we used recombinant murine EGF, murine platelet-derived growth factor bb (PDGFbb), human EGF, human vascular endothelial growth factor (VEGF), murine M-CSF, and murine RANKL (Peprotech) reconstituted in water. .. Seven micrometer sections of synovial tissue from patients with RA were fixed in 4% (v/v) paraformeldehyde, permeabilized with triton X-100, and probed with antibodies against total EGFR (Clone LA-1, Millipore) or EGFR phosphorylated at tyrosine 1068 (p-EGFR(Y1068); Abcam).

    Article Title: Reversible Block of Mouse Neural Stem Cell Differentiation in the Absence of Dicer and MicroRNAs
    Article Snippet: Neural stem cell line derivation, culture and characterization Polyclonal neural stem cell lines were established from the E12.5 cortex of Dicer-null and wild-type littermate mice and maintained according to published methods with minor variations . .. Briefly, E12.5 cortices were dissociated to single cells and placed in non-adherent culture conditions to promote neurosphere formation in the presence of 10 µg/ml recombinant EGF and 10 µg/ml recombinant FGF2 (Peprotech). ..

    Imaging:

    Article Title: Long chain n-3 fatty acids attenuate oncogenic KRas-driven proliferation by altering plasma membrane nanoscale proteolipid composition
    Article Snippet: Confocal images were collected using a Nikon Eclipse Ti confocal system and processed using Nikon software and Adobe Photoshop. .. For quantitative phospho-ERK analysis, cells were seeded in cell imaging 8 chamber coverglass slides (Eppendorf, 0030742036) and treated with select fatty acids (50 μM) for 72 h. Cells were subsequently serum starved (0% FBS) for 18 h in the presence of fatty acid, stimulated with EGF (25 ng/ml) (PeproTech, 315-09) for 5 min and immediately fixed in ice cold 100% methanol. .. Cells were incubated with primary phosphor-ERK (Cell Signaling Technology, #4370, 1:200, RRID:AB_2315112) antibody followed by alexa-647 fluorescent secondary antibody (Jackson ImmunoResearch Labs, #711-605-152, 1:400, RRID:AB_2492288).

    Modification:

    Article Title: Self-renewing Monolayer of Primary Colonic or Rectal Epithelial Cells
    Article Snippet: .. For mouse cells, the culture medium (ENR-W) was advanced Dulbecco modified Eagle medium/F12 medium with Wnt-3A (30 ng/mL), R-spondin-2 (75 ng/mL), noggin (71 ng/mL), EGF (50 ng/mL; PeproTech 315-09, Rocky Hill, NJ), N-acetyl cysteine (1 mmol/L, Sigma A9165; Sigma-Aldrich), GlutaMAX (×1), HEPES (10 mmol/L), A83-01 (500 ng/mL, Sigma SML0788), penicillin (100 units/mL), streptomycin (100 μg/mL), and gentamicin (5 μg/mL). .. For human cells, the human intestinal stem cell medium (HISC) was same as ENR-W, except that Wnt-3A (100 ng/mL), R-spondin-2 (90 ng/mL), N-acetylcysteine (1.25 mmol/L) and additional B27 (×1), nicotinamide (10 mmol/L, Sigma N0636), gastrin (10 nmol/L, AnaSpec AS-64149; Fremont, CA), SB202190 (3 μmol/L, Selleckchem S1077; Houston, TX), and prostaglandin E2 (10 nmol/L, Cayman Chemicals 14010; Ann Arbor, MI) were added.

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    • h egf  (PeproTech)
    99
    PeproTech h egf
    Schematic representations of <t>EGF-</t> and <t>MIF-induced</t> activation of P-ERK1, 2 and the phosphorylation of MSK1/2 and RSK1/2 kinases. P-MSK1/2 induces P-NF-κB/p65 (S276) and the transcription of PP2Cδ/Wip1, which is inhibited by DMF; 24 hours stimulation by EGF and MIF induces P-Wip1/Wip1, resulting in dephosphorylation of P-RSK2 (S386) in complex formation with P-RSK2. This is inhibited by costimulation with the ERK1/2 inhibitor PD98059 or DMF. Abbreviations: DMF, dimethyl fumarate; EGF, epidermal growth factor; ERK, extracellular signal-regulated kinase; MIF, macrophage migration inhibitory factor; MSK, mitogen- and stress-activated kinase; NF-κB, nuclear factor-κB; PP2Cδ, protein phosphatase 2Cδ; RSK, p90 ribosomal S6 kinase; Wip1, wild-type p53-induced phosphatase 1.
    H Egf, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/h egf/product/PeproTech
    Average 99 stars, based on 1 article reviews
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    egf  (PeproTech)
    99
    PeproTech egf
    Synergistic effect of <t>PDGFaa</t> and <t>EGF</t> in promoting the self-renewal of O4 − OPCs. (A) Representative images of O4 + and/or BrdU + cells cultured in PDGFaa or PDGFaa + EGF for 5 days, cell proliferation was analyzed by BrdU incorporation for 24 h before fixation. (B) Quantification of O4 + cells in PDGFaa and PDGFaa + EGF cultures. (C) Quantification of BrdU + cells in PDGFaa and PDGFaa + EGF cultures. Statistical analyses are presented as mean ± SD, n = 3. * P
    Egf, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    egf - by Bioz Stars, 2021-04
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    human egf  (PeproTech)
    86
    PeproTech human egf
    Changes in R b as a consequence of altering growth medium components as shown by brain endothelium. Endothelial cells were seeded at 20,000 cells per well on a 96w20idf ECIS array. Time 0 h denotes the time cells were seeded. The dotted vertical line indicates 48 h of cell growth in each respective media type, with a subsequent media change carried out at this time. ( A ) Brain endothelium grown in Enriched Media containing either 10%, 5%, or 2% FBS; ( B ) Brain endothelium grown in Enriched Media containing either 160 µM, 80 µM, 40 µM, or 0 µM <t>cAMP;</t> ( C ) Brain endothelium grown in Enriched Media containing either 1 µg/mL, 39 ng/mL, or 0 ng/mL hydrocortisone; ( D ) Brain endothelium grown in Enriched Media containing either 1 ng/mL or 0 ng/mL <t>EGF;</t> ( E ) Brain endothelium grown in Enriched Media containing either 3 ng/mL or 0 ng/mL FGF; ( F ) Brain endothelium grown in Enriched Media containing either 10 µg/mL or 0 µg/mL heparin; ( G ) Brain endothelium grown in Enriched Media containing either 1× or no Glutamax. Data show the mean ± SD (n = 3 wells) of one independent experiment representative of three experimental repeats. Graphical representations of p values are * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.
    Human Egf, supplied by PeproTech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hb egf  (PeproTech)
    95
    PeproTech hb egf
    Impaired <t>EGFR</t> signaling pathway in ZBTB20-deficient hepatocytes can be corrected by EGFR overexpression. a Primary hepatocytes from LZB20KO and control mice were stimulated with <t>EGF</t> (1 ng/ml) for indicated times following 12 h of serum starvation, and the expression and phosphorylation of EGFR, ERK, and AKT were analyzed by western blot. b Hepatocytes were infected with adenoviruses Ad-EGFR or Ad-GFP (MOI = 10) at 24 h prior to 12 h of serum starvation, and followed by EGF stimulation for 5 min. The expression and phosphorylation of EGFR, ERK, and AKT were analyzed by western blot
    Hb Egf, supplied by PeproTech, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Schematic representations of EGF- and MIF-induced activation of P-ERK1, 2 and the phosphorylation of MSK1/2 and RSK1/2 kinases. P-MSK1/2 induces P-NF-κB/p65 (S276) and the transcription of PP2Cδ/Wip1, which is inhibited by DMF; 24 hours stimulation by EGF and MIF induces P-Wip1/Wip1, resulting in dephosphorylation of P-RSK2 (S386) in complex formation with P-RSK2. This is inhibited by costimulation with the ERK1/2 inhibitor PD98059 or DMF. Abbreviations: DMF, dimethyl fumarate; EGF, epidermal growth factor; ERK, extracellular signal-regulated kinase; MIF, macrophage migration inhibitory factor; MSK, mitogen- and stress-activated kinase; NF-κB, nuclear factor-κB; PP2Cδ, protein phosphatase 2Cδ; RSK, p90 ribosomal S6 kinase; Wip1, wild-type p53-induced phosphatase 1.

    Journal: Journal of Inflammation Research

    Article Title: Protein phosphatase 2Cδ/Wip1 regulates phospho-p90RSK2 activity in lesional psoriatic skin

    doi: 10.2147/JIR.S152869

    Figure Lengend Snippet: Schematic representations of EGF- and MIF-induced activation of P-ERK1, 2 and the phosphorylation of MSK1/2 and RSK1/2 kinases. P-MSK1/2 induces P-NF-κB/p65 (S276) and the transcription of PP2Cδ/Wip1, which is inhibited by DMF; 24 hours stimulation by EGF and MIF induces P-Wip1/Wip1, resulting in dephosphorylation of P-RSK2 (S386) in complex formation with P-RSK2. This is inhibited by costimulation with the ERK1/2 inhibitor PD98059 or DMF. Abbreviations: DMF, dimethyl fumarate; EGF, epidermal growth factor; ERK, extracellular signal-regulated kinase; MIF, macrophage migration inhibitory factor; MSK, mitogen- and stress-activated kinase; NF-κB, nuclear factor-κB; PP2Cδ, protein phosphatase 2Cδ; RSK, p90 ribosomal S6 kinase; Wip1, wild-type p53-induced phosphatase 1.

    Article Snippet: After 24 hours, the cells were either left alone or preincubated with DMF (140 µM) or 50 µM of PD98059 for 1 hour and stimulated with recombinant MIF (100 ng/mL; R & D Systems, Oxon, UK) or h-EGF (2 ng/mL; PeproTech, London, UK) for 24 hours.

    Techniques: Activation Assay, De-Phosphorylation Assay, Migration

    ( A ) Complex formation of Wip1 with RSK2 in human keratinocytes was induced by EGF and MIF. Keratinocytes were either left alone or preincubated with DMF (140 µM) or PD98059 (50 µM) for 1 hour and stimulated with EGF (2 ng/mL) or MIF (100 ng/mL) for 24 hours. Whole cell extracts were immunoprecipitated with anti-RSK2 antibody and analyzed by WB with anti-Wip1 (H-300) (sc-20712 epitope C-terminus) and anti-RSK2; 1 representative gel is shown. ( B ) Densitometry of 6 different experiments from pooled data shows the ratio of P-Wip1/Wip1 (both) associated with RSK2. EGF-stimulated versus (co) untreated cells * p =0.003; MIF stimulated versus (co) untreated cells ¤ p =0.037; EGF-stimulated versus (DMF and EGF) ** p =0.008; MIF-stimulated versus (DMF and MIF) p =0.059; EGF-stimulated versus (PD98059 and EGF) *** p

    Journal: Journal of Inflammation Research

    Article Title: Protein phosphatase 2Cδ/Wip1 regulates phospho-p90RSK2 activity in lesional psoriatic skin

    doi: 10.2147/JIR.S152869

    Figure Lengend Snippet: ( A ) Complex formation of Wip1 with RSK2 in human keratinocytes was induced by EGF and MIF. Keratinocytes were either left alone or preincubated with DMF (140 µM) or PD98059 (50 µM) for 1 hour and stimulated with EGF (2 ng/mL) or MIF (100 ng/mL) for 24 hours. Whole cell extracts were immunoprecipitated with anti-RSK2 antibody and analyzed by WB with anti-Wip1 (H-300) (sc-20712 epitope C-terminus) and anti-RSK2; 1 representative gel is shown. ( B ) Densitometry of 6 different experiments from pooled data shows the ratio of P-Wip1/Wip1 (both) associated with RSK2. EGF-stimulated versus (co) untreated cells * p =0.003; MIF stimulated versus (co) untreated cells ¤ p =0.037; EGF-stimulated versus (DMF and EGF) ** p =0.008; MIF-stimulated versus (DMF and MIF) p =0.059; EGF-stimulated versus (PD98059 and EGF) *** p

    Article Snippet: After 24 hours, the cells were either left alone or preincubated with DMF (140 µM) or 50 µM of PD98059 for 1 hour and stimulated with recombinant MIF (100 ng/mL; R & D Systems, Oxon, UK) or h-EGF (2 ng/mL; PeproTech, London, UK) for 24 hours.

    Techniques: Immunoprecipitation, Western Blot

    Synergistic effect of PDGFaa and EGF in promoting the self-renewal of O4 − OPCs. (A) Representative images of O4 + and/or BrdU + cells cultured in PDGFaa or PDGFaa + EGF for 5 days, cell proliferation was analyzed by BrdU incorporation for 24 h before fixation. (B) Quantification of O4 + cells in PDGFaa and PDGFaa + EGF cultures. (C) Quantification of BrdU + cells in PDGFaa and PDGFaa + EGF cultures. Statistical analyses are presented as mean ± SD, n = 3. * P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: EGF Enhances Oligodendrogenesis from Glial Progenitor Cells

    doi: 10.3389/fnmol.2017.00106

    Figure Lengend Snippet: Synergistic effect of PDGFaa and EGF in promoting the self-renewal of O4 − OPCs. (A) Representative images of O4 + and/or BrdU + cells cultured in PDGFaa or PDGFaa + EGF for 5 days, cell proliferation was analyzed by BrdU incorporation for 24 h before fixation. (B) Quantification of O4 + cells in PDGFaa and PDGFaa + EGF cultures. (C) Quantification of BrdU + cells in PDGFaa and PDGFaa + EGF cultures. Statistical analyses are presented as mean ± SD, n = 3. * P

    Article Snippet: A2B5+ cells were then grown in glial basal medium (DMEM/F12, 1 × N2, 1 × B27, 1 × P/S, and 0.1% w/v BSA, all from Gibco) supplemented with 10 ng/ml PDGFaa and EGF (Peprotech) on fibronectin/laminin coated 12-well plates at 2000 cells/well for mass culture experiments or on coated 24-well plated at 1000 cells/well for immunofluorescence staining.

    Techniques: Cell Culture, BrdU Incorporation Assay

    Biological effects of EGF, PDGFaa, bFGF and T3 in the progression of OL lineage. EGF has multiple biological effects on oligodendrogenesis, and its functional output is influenced by other signal molecules, such as PDGFaa and T3. In GRP cells, EGF and bFGF collaborate with PDGFaa to promote the self-renewal of GRP cells. When EGF is present alone, it favors the development of GRP cells to OPCs, and this progress is accelerated by supplementing PDGFaa and T3 simultaneously. In OPCs, EGF and bFGF enhance their responsiveness to PDGFaa and thus maintains their O4 negative phenotype as well as bipolar or tripolar cell morphology. When EGF is present alone, it synergizes with T3 to promote the terminal differentiation of OPCs, whereas bFGF and PDGFaa inhibit this differentiation process.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: EGF Enhances Oligodendrogenesis from Glial Progenitor Cells

    doi: 10.3389/fnmol.2017.00106

    Figure Lengend Snippet: Biological effects of EGF, PDGFaa, bFGF and T3 in the progression of OL lineage. EGF has multiple biological effects on oligodendrogenesis, and its functional output is influenced by other signal molecules, such as PDGFaa and T3. In GRP cells, EGF and bFGF collaborate with PDGFaa to promote the self-renewal of GRP cells. When EGF is present alone, it favors the development of GRP cells to OPCs, and this progress is accelerated by supplementing PDGFaa and T3 simultaneously. In OPCs, EGF and bFGF enhance their responsiveness to PDGFaa and thus maintains their O4 negative phenotype as well as bipolar or tripolar cell morphology. When EGF is present alone, it synergizes with T3 to promote the terminal differentiation of OPCs, whereas bFGF and PDGFaa inhibit this differentiation process.

    Article Snippet: A2B5+ cells were then grown in glial basal medium (DMEM/F12, 1 × N2, 1 × B27, 1 × P/S, and 0.1% w/v BSA, all from Gibco) supplemented with 10 ng/ml PDGFaa and EGF (Peprotech) on fibronectin/laminin coated 12-well plates at 2000 cells/well for mass culture experiments or on coated 24-well plated at 1000 cells/well for immunofluorescence staining.

    Techniques: Functional Assay

    Expanded clones of A2B5 + cell expressed typical differentiation phenotype of tripotential glial restricted precursor (GRP) cells. (A) An example of an expanded clone in presence of EGF + PDGFaa. (B) A2B5 + clones were digested and replated into separate wells of 24-well plates and induced to differentiate for 5 days in the presence of bFGF + CNTF or FBS, A2B5 + /GFAP + and A2B5 − /GFAP + astrocytes were obtained, respectively. (C) Quantification of A2B5 + /GFAP + and A2B5 − /GFAP + astrocytes in the differentiation cultures exposed to bFGF + CNTF and FBS, n = 3. (D) A2B5 + cells were cultured in T3 for 5 days and MBP + Oligodendrocytes (OLs) can be detected. (E) The culture described in (D) was confirmed by western blotting with MBP antibody. Abbreviation: OL, oligodendrocyte. ** P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: EGF Enhances Oligodendrogenesis from Glial Progenitor Cells

    doi: 10.3389/fnmol.2017.00106

    Figure Lengend Snippet: Expanded clones of A2B5 + cell expressed typical differentiation phenotype of tripotential glial restricted precursor (GRP) cells. (A) An example of an expanded clone in presence of EGF + PDGFaa. (B) A2B5 + clones were digested and replated into separate wells of 24-well plates and induced to differentiate for 5 days in the presence of bFGF + CNTF or FBS, A2B5 + /GFAP + and A2B5 − /GFAP + astrocytes were obtained, respectively. (C) Quantification of A2B5 + /GFAP + and A2B5 − /GFAP + astrocytes in the differentiation cultures exposed to bFGF + CNTF and FBS, n = 3. (D) A2B5 + cells were cultured in T3 for 5 days and MBP + Oligodendrocytes (OLs) can be detected. (E) The culture described in (D) was confirmed by western blotting with MBP antibody. Abbreviation: OL, oligodendrocyte. ** P

    Article Snippet: A2B5+ cells were then grown in glial basal medium (DMEM/F12, 1 × N2, 1 × B27, 1 × P/S, and 0.1% w/v BSA, all from Gibco) supplemented with 10 ng/ml PDGFaa and EGF (Peprotech) on fibronectin/laminin coated 12-well plates at 2000 cells/well for mass culture experiments or on coated 24-well plated at 1000 cells/well for immunofluorescence staining.

    Techniques: Clone Assay, Cell Culture, Western Blot

    EGF enhanced the formation of OPCs from GRP clones. (A) The single cell tracking of a GRP cell generating O4 + /MBP − daughter cells in EGF for 5 days. With the progression of cell division, a fibroblast-like GRP cell was gradually converted to typical OPCs of bipolar or tripolar morphology, well separated from each other, and expressed O4 antigen but not MBP. Daughter cells of the same progenitor were indicated by arrows of different color. (B) Quantification of the clones containing O4 + /MBP − cells in the GRP cell cultures exposed to various combinations of factors for 5 days, respectively, n = 3. (C) OPC-like cells from (B) differentiated into MBP + cells after 5 days of culture in glial basal medium without supplemented factors. Left: phase image; right: anti-MBP immunostaining. (D) Quantification of the clones containing MBP + cells in OL differentiation cultures described in (C) , n = 3. (E) OPCs derived from EGF + PDGFaa + T3 treament differentiated into A2B5 + /GFAP + astrocytes in presence of FBS. * P

    Journal: Frontiers in Molecular Neuroscience

    Article Title: EGF Enhances Oligodendrogenesis from Glial Progenitor Cells

    doi: 10.3389/fnmol.2017.00106

    Figure Lengend Snippet: EGF enhanced the formation of OPCs from GRP clones. (A) The single cell tracking of a GRP cell generating O4 + /MBP − daughter cells in EGF for 5 days. With the progression of cell division, a fibroblast-like GRP cell was gradually converted to typical OPCs of bipolar or tripolar morphology, well separated from each other, and expressed O4 antigen but not MBP. Daughter cells of the same progenitor were indicated by arrows of different color. (B) Quantification of the clones containing O4 + /MBP − cells in the GRP cell cultures exposed to various combinations of factors for 5 days, respectively, n = 3. (C) OPC-like cells from (B) differentiated into MBP + cells after 5 days of culture in glial basal medium without supplemented factors. Left: phase image; right: anti-MBP immunostaining. (D) Quantification of the clones containing MBP + cells in OL differentiation cultures described in (C) , n = 3. (E) OPCs derived from EGF + PDGFaa + T3 treament differentiated into A2B5 + /GFAP + astrocytes in presence of FBS. * P

    Article Snippet: A2B5+ cells were then grown in glial basal medium (DMEM/F12, 1 × N2, 1 × B27, 1 × P/S, and 0.1% w/v BSA, all from Gibco) supplemented with 10 ng/ml PDGFaa and EGF (Peprotech) on fibronectin/laminin coated 12-well plates at 2000 cells/well for mass culture experiments or on coated 24-well plated at 1000 cells/well for immunofluorescence staining.

    Techniques: Single Cell Tracking, Clone Assay, Immunostaining, Derivative Assay

    Changes in R b as a consequence of altering growth medium components as shown by brain endothelium. Endothelial cells were seeded at 20,000 cells per well on a 96w20idf ECIS array. Time 0 h denotes the time cells were seeded. The dotted vertical line indicates 48 h of cell growth in each respective media type, with a subsequent media change carried out at this time. ( A ) Brain endothelium grown in Enriched Media containing either 10%, 5%, or 2% FBS; ( B ) Brain endothelium grown in Enriched Media containing either 160 µM, 80 µM, 40 µM, or 0 µM cAMP; ( C ) Brain endothelium grown in Enriched Media containing either 1 µg/mL, 39 ng/mL, or 0 ng/mL hydrocortisone; ( D ) Brain endothelium grown in Enriched Media containing either 1 ng/mL or 0 ng/mL EGF; ( E ) Brain endothelium grown in Enriched Media containing either 3 ng/mL or 0 ng/mL FGF; ( F ) Brain endothelium grown in Enriched Media containing either 10 µg/mL or 0 µg/mL heparin; ( G ) Brain endothelium grown in Enriched Media containing either 1× or no Glutamax. Data show the mean ± SD (n = 3 wells) of one independent experiment representative of three experimental repeats. Graphical representations of p values are * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Journal: Biosensors

    Article Title: The Importance of Multifrequency Impedance Sensing of Endothelial Barrier Formation Using ECIS Technology for the Generation of a Strong and Durable Paracellular Barrier

    doi: 10.3390/bios8030064

    Figure Lengend Snippet: Changes in R b as a consequence of altering growth medium components as shown by brain endothelium. Endothelial cells were seeded at 20,000 cells per well on a 96w20idf ECIS array. Time 0 h denotes the time cells were seeded. The dotted vertical line indicates 48 h of cell growth in each respective media type, with a subsequent media change carried out at this time. ( A ) Brain endothelium grown in Enriched Media containing either 10%, 5%, or 2% FBS; ( B ) Brain endothelium grown in Enriched Media containing either 160 µM, 80 µM, 40 µM, or 0 µM cAMP; ( C ) Brain endothelium grown in Enriched Media containing either 1 µg/mL, 39 ng/mL, or 0 ng/mL hydrocortisone; ( D ) Brain endothelium grown in Enriched Media containing either 1 ng/mL or 0 ng/mL EGF; ( E ) Brain endothelium grown in Enriched Media containing either 3 ng/mL or 0 ng/mL FGF; ( F ) Brain endothelium grown in Enriched Media containing either 10 µg/mL or 0 µg/mL heparin; ( G ) Brain endothelium grown in Enriched Media containing either 1× or no Glutamax. Data show the mean ± SD (n = 3 wells) of one independent experiment representative of three experimental repeats. Graphical representations of p values are * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

    Article Snippet: Cells were maintained in M199 media (Gibco, Waltham, MA, USA) supplemented with 10% FBS, 1 µg/mL hydrocortisone (Sigma, St Louis, MO, USA), 80 µM butyryl cAMP (Sigma, St Louis, MO, USA), 10 µg/mL heparin (Sigma), 1 ng/mL human EGF (Peprotech, Rocky Hill, NJ, USA), 3 ng/mL human FGF (Peprotech, Rocky Hill, NJ, USA), and 2 mM Glutamax (Gibco, Waltham, MA, USA).

    Techniques: Electric Cell-substrate Impedance Sensing

    Impaired EGFR signaling pathway in ZBTB20-deficient hepatocytes can be corrected by EGFR overexpression. a Primary hepatocytes from LZB20KO and control mice were stimulated with EGF (1 ng/ml) for indicated times following 12 h of serum starvation, and the expression and phosphorylation of EGFR, ERK, and AKT were analyzed by western blot. b Hepatocytes were infected with adenoviruses Ad-EGFR or Ad-GFP (MOI = 10) at 24 h prior to 12 h of serum starvation, and followed by EGF stimulation for 5 min. The expression and phosphorylation of EGFR, ERK, and AKT were analyzed by western blot

    Journal: Cell Death & Disease

    Article Title: ZBTB20 regulates EGFR expression and hepatocyte proliferation in mouse liver regeneration

    doi: 10.1038/s41419-018-0514-0

    Figure Lengend Snippet: Impaired EGFR signaling pathway in ZBTB20-deficient hepatocytes can be corrected by EGFR overexpression. a Primary hepatocytes from LZB20KO and control mice were stimulated with EGF (1 ng/ml) for indicated times following 12 h of serum starvation, and the expression and phosphorylation of EGFR, ERK, and AKT were analyzed by western blot. b Hepatocytes were infected with adenoviruses Ad-EGFR or Ad-GFP (MOI = 10) at 24 h prior to 12 h of serum starvation, and followed by EGF stimulation for 5 min. The expression and phosphorylation of EGFR, ERK, and AKT were analyzed by western blot

    Article Snippet: Hepatocytes were stimulated by 20 ng/ml HGF (Peprotech), 10 ng/ml TNF-α (Peprotech), or EGFR ligands such as 1 ng/ml EGF (Peprotech), 1 ng/ml HB-EGF (Peprotech), and 10 ng/ml amphiregulin (Peprotech) for various minutes after serum starvation of 12 h. For adenovirus-mediated gene transfection, hepatocytes were cultured in the presence of adenoviruses with the multiplicity of infection (MOI) of 100 for 6 h, and refreshed with normal media later.

    Techniques: Over Expression, Mouse Assay, Expressing, Western Blot, Infection