human erk1 2  (Cell Signaling Technology Inc)

 
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    Name:
    p44 42 MAPK Erk1 2 3A7 Mouse mAb
    Description:
    Mitogen activated protein kinases MAPKs are a widely conserved family of serine threonine protein kinases involved in many cellular programs such as cell proliferation differentiation motility and death The p44 42 MAPK Erk1 2 signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens growth factors and cytokines 1 3 and research investigators consider it an important target in the diagnosis and treatment of cancer 4 Upon stimulation a sequential three part protein kinase cascade is initiated consisting of a MAP kinase kinase kinase MAPKKK or MAP3K a MAP kinase kinase MAPKK or MAP2K and a MAP kinase MAPK Multiple p44 42 MAP3Ks have been identified including members of the Raf family as well as Mos and Tpl2 COT MEK1 and MEK2 are the primary MAPKKs in this pathway 5 6 MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202 Tyr204 and Thr185 Tyr187 respectively Several downstream targets of p44 42 have been identified including p90RSK 7 and the transcription factor Elk 1 8 9 p44 42 are negatively regulated by a family of dual specificity Thr Tyr MAPK phosphatases known as DUSPs or MKPs 10 along with MEK inhibitors such as U0126 and PD98059
    Catalog Number:
    9107
    Price:
    None
    Category:
    Primary Antibodies
    Source:
    Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of p44/42 MAP kinase.
    Reactivity:
    Human Mouse Rat Hamster Monkey Mink Zebrafish Bovine Pig
    Applications:
    Western Blot
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    Structured Review

    Cell Signaling Technology Inc human erk1 2
    IL-2 augmented phosphorylation/activation of cellular <t>ERK1/2.</t> (a) Western blotting showed that the presence of IL-2 (40 units/ml) in cultures significantly augmented the activation of cellular p-ERK1/2 in Karpas 299 and SUDHL-1 cells but exerted no effect on the protein expression of NPM-ALK as an indicator of equal protein loading. (b) Exposure of Karpas 299 cells to the kinase inhibitor U0126 (5 μM) resulted in complete inhibition of the constitutive ERK1/2 phosphorylation (lanes 1 and 2 of upper panel). IL-2 treatment overcame this U0126-induced decrease of ERK1/2 phosphorylation (lanes 2 and 4 of upper panel). Total ERK1/2 protein levels showed no change in the presence of U0126 and/or IL-2 treatments (lower panel). (c) The similar results were also observed with SUDHL-1 cells under the same treatments. The results are representative of three separate experiments with similar findings.
    Mitogen activated protein kinases MAPKs are a widely conserved family of serine threonine protein kinases involved in many cellular programs such as cell proliferation differentiation motility and death The p44 42 MAPK Erk1 2 signaling pathway can be activated in response to a diverse range of extracellular stimuli including mitogens growth factors and cytokines 1 3 and research investigators consider it an important target in the diagnosis and treatment of cancer 4 Upon stimulation a sequential three part protein kinase cascade is initiated consisting of a MAP kinase kinase kinase MAPKKK or MAP3K a MAP kinase kinase MAPKK or MAP2K and a MAP kinase MAPK Multiple p44 42 MAP3Ks have been identified including members of the Raf family as well as Mos and Tpl2 COT MEK1 and MEK2 are the primary MAPKKs in this pathway 5 6 MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202 Tyr204 and Thr185 Tyr187 respectively Several downstream targets of p44 42 have been identified including p90RSK 7 and the transcription factor Elk 1 8 9 p44 42 are negatively regulated by a family of dual specificity Thr Tyr MAPK phosphatases known as DUSPs or MKPs 10 along with MEK inhibitors such as U0126 and PD98059
    https://www.bioz.com/result/human erk1 2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human erk1 2 - by Bioz Stars, 2021-06
    94/100 stars

    Images

    1) Product Images from "Interleukin-2 Functions in Anaplastic Large Cell Lymphoma Cells through Augmentation of Extracellular Signal-Regulated Kinases 1/2 Activation"

    Article Title: Interleukin-2 Functions in Anaplastic Large Cell Lymphoma Cells through Augmentation of Extracellular Signal-Regulated Kinases 1/2 Activation

    Journal: International Journal of Biomedical Science : IJBS

    doi:

    IL-2 augmented phosphorylation/activation of cellular ERK1/2. (a) Western blotting showed that the presence of IL-2 (40 units/ml) in cultures significantly augmented the activation of cellular p-ERK1/2 in Karpas 299 and SUDHL-1 cells but exerted no effect on the protein expression of NPM-ALK as an indicator of equal protein loading. (b) Exposure of Karpas 299 cells to the kinase inhibitor U0126 (5 μM) resulted in complete inhibition of the constitutive ERK1/2 phosphorylation (lanes 1 and 2 of upper panel). IL-2 treatment overcame this U0126-induced decrease of ERK1/2 phosphorylation (lanes 2 and 4 of upper panel). Total ERK1/2 protein levels showed no change in the presence of U0126 and/or IL-2 treatments (lower panel). (c) The similar results were also observed with SUDHL-1 cells under the same treatments. The results are representative of three separate experiments with similar findings.
    Figure Legend Snippet: IL-2 augmented phosphorylation/activation of cellular ERK1/2. (a) Western blotting showed that the presence of IL-2 (40 units/ml) in cultures significantly augmented the activation of cellular p-ERK1/2 in Karpas 299 and SUDHL-1 cells but exerted no effect on the protein expression of NPM-ALK as an indicator of equal protein loading. (b) Exposure of Karpas 299 cells to the kinase inhibitor U0126 (5 μM) resulted in complete inhibition of the constitutive ERK1/2 phosphorylation (lanes 1 and 2 of upper panel). IL-2 treatment overcame this U0126-induced decrease of ERK1/2 phosphorylation (lanes 2 and 4 of upper panel). Total ERK1/2 protein levels showed no change in the presence of U0126 and/or IL-2 treatments (lower panel). (c) The similar results were also observed with SUDHL-1 cells under the same treatments. The results are representative of three separate experiments with similar findings.

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

    IL-2 regulated ALCL cell growth and apoptosis. To downregulate the ERK1/2 pathway, cultured Karpas 299 (a) and SUDHL-1 cells (b) were exposed to U0126 (5 μM) in the presence or absence of IL-2 (40 units/ml) as indicated. After culture for 36 hours, the number of viable cells in each condition was counted. The presence of exogenous IL-2 significantly reversed the U0126-induced growth arrest of ALCL cells. The treated cells were also stained with FITC-conjugated annexin V, and the apoptotic rate (%) was determined by flow cytometry. The presence of exogenous IL-2 almost completely suppressed the U0126-induced apoptosis of Karpas 299 (c) and SUDHL-1 cells (d). * p
    Figure Legend Snippet: IL-2 regulated ALCL cell growth and apoptosis. To downregulate the ERK1/2 pathway, cultured Karpas 299 (a) and SUDHL-1 cells (b) were exposed to U0126 (5 μM) in the presence or absence of IL-2 (40 units/ml) as indicated. After culture for 36 hours, the number of viable cells in each condition was counted. The presence of exogenous IL-2 significantly reversed the U0126-induced growth arrest of ALCL cells. The treated cells were also stained with FITC-conjugated annexin V, and the apoptotic rate (%) was determined by flow cytometry. The presence of exogenous IL-2 almost completely suppressed the U0126-induced apoptosis of Karpas 299 (c) and SUDHL-1 cells (d). * p

    Techniques Used: Cell Culture, Staining, Flow Cytometry, Cytometry

    Related Articles

    Western Blot:

    Article Title: Deletion of Na+/H+ exchanger regulatory factor 2 represses colon cancer progress by suppression of Stat3 and CD24
    Article Snippet: Additional rabbit polyclonal anti-NHERF2 (HPA001672, 1:250 dilution for IHC) and mouse monoclonal anti-β-actin (A1978, 1:5,000 for Western blot) antibodies were obtained from Sigma. .. Rabbit monoclonal anti-phospho-ERK1/2 (p-ERK1/2) (no. 4370, 1:2,000 for Western blot and 1:400 for IHC), mouse monoclonal anti-ERK1/2 (no. 4696, 1:2,000 for Western blot), rabbit monoclonal anti-phospho-Stat3 (no. 9145, 1:2,000 for Western blot and 1:400 for IHC), and rabbit monoclonal anti-Stat3 (no. 4904, 1:2,000 for Western blot) antibodies were purchased from Cell Signaling. .. Statistical significance was assessed by two-tailed unpaired Student's t- test, Welch's t -test, Mann-Whitney's U -test, or mxn χ2 test.

    Article Title: Targeting Long Noncoding RNA HMMR-AS1 Suppresses and Radiosensitizes Glioblastoma
    Article Snippet: The luciferin (in vivo grade) used for animal experiments was purchased from Promega (Madison, WI). .. For Western blot analysis, the primary antibodies against β-actin, c-Myc, BMI1, p27 Kip1, Cyclin D1, CDK2, CDK4, ERK1/2, p-ERK1/2(Thr202/Tyr204), ZEB1, β-Catenin, N-Cadherin, Vimentin, ATM, p-ATM, and Rad51 were all purchased from Cell Signaling Technology (Beverly, MA); HMMR#1 was purchased from OriGene (Rockville, MD); and HMMR#2 was purchased from GeneTex (Irvine, CA). .. For immunohistochemical (IHC) staining, the primary antibody HMMR#1 was purchased from Origene (Rockville, MD), ZEB1 was from Abcam (Cambridge, MA), and β-Catenin and Vimentin were purchased from Cell Signaling Technology (Beverly, MA).

    Article Title: Interleukin-2 Functions in Anaplastic Large Cell Lymphoma Cells through Augmentation of Extracellular Signal-Regulated Kinases 1/2 Activation
    Article Snippet: Apoptotic cells (%) were detected by flow cytometric analysis. .. Western blotting of cellular protein kinases Antibodies for human ERK1/2, phosphorylated ERK1/2, and NPM-ALK proteins were purchased from Cell Signaling (Beverly, MA). .. Western blotting was performed on extracts from Karpas 299 and SUDHL-1 cells (5 × 106 cells/sample) treated with IL-2 (40 units/ml) and/or exposed to U0126 (5 μM) for 30 minutes.

    Immunohistochemistry:

    Article Title: Deletion of Na+/H+ exchanger regulatory factor 2 represses colon cancer progress by suppression of Stat3 and CD24
    Article Snippet: Additional rabbit polyclonal anti-NHERF2 (HPA001672, 1:250 dilution for IHC) and mouse monoclonal anti-β-actin (A1978, 1:5,000 for Western blot) antibodies were obtained from Sigma. .. Rabbit monoclonal anti-phospho-ERK1/2 (p-ERK1/2) (no. 4370, 1:2,000 for Western blot and 1:400 for IHC), mouse monoclonal anti-ERK1/2 (no. 4696, 1:2,000 for Western blot), rabbit monoclonal anti-phospho-Stat3 (no. 9145, 1:2,000 for Western blot and 1:400 for IHC), and rabbit monoclonal anti-Stat3 (no. 4904, 1:2,000 for Western blot) antibodies were purchased from Cell Signaling. .. Statistical significance was assessed by two-tailed unpaired Student's t- test, Welch's t -test, Mann-Whitney's U -test, or mxn χ2 test.

    Blocking Assay:

    Article Title: Scratching Counteracts IL-13 Signaling by Upregulating the Decoy Receptor IL-13Rα2 in Keratinocytes
    Article Snippet: Equal 20 μg amounts of protein were mixed with 4× LDS sample buffer (Invitrogen) and 10× sample reducing agent (Invitrogen), boiled at 70 °C for 10 min, loaded onto Bolt 4–12% Bis-Tris Plus (Thermo Fisher Scientific), and electrophoresed using Power Station III (Atto corporation, Tokyo, Japan) at 200 V and 180 mA for 25 min. .. The proteins were then transferred to an Immobilon PVDF Transfer Membrane (Merck, Kenilworth, NJ, USA), using Power Station III at 30 V for 1 h. Membranes were blocked with a blocking buffer, containing blocker diluent A and B, (Invitrogen) for 30 min. Membranes were probed overnight at 4 °C with the following primary antibodies: β-actin (Cell Signaling Technology), ERK1/2, JNK, p38, Phospho-ERK1/2, Phospho-JNK, and Phospho-p38 (Cell Signaling Technology). .. The secondary antibodies, anti-mouse IgG HRP-linked antibody (Cell Signaling Technology) and anti-rabbit IgG HRP-linked antibody (Cell Signaling Technology), were applied at room temperature for 30 min. Visualization of protein bands was accomplished with Super Signal West Pico Chemiluminescent Substrate (Thermo Fisher Scientific), using the ChemiDoc Touch Imaging System (Bio-Rad).

    Article Title: FGF21 promotes ischaemic angiogenesis and endothelial progenitor cells function under diabetic conditions in an AMPK/NAD+‐dependent manner, et al. FGF21 promotes ischaemic angiogenesis and endothelial progenitor cells function under diabetic conditions in an AMPK/NAD+‐dependent manner
    Article Snippet: After determining the protein concentration with Quick Start Bradford Dye Reagent (Bio‐Rad), proteins were separated on a 10% SDS‐PAGE and transferred onto nitrocellulose membranes (Bio‐Rad). .. After blocking in 5% skim milk for 1 hour, the membranes were incubated overnight at 4°C with antibodies against phospho‐extracellular regulated protein kinases 1/2 (p‐Erk1/2), Erk1/2, CD38, NAMPT, poly (ADP‐ribose) polymerase (PARP), CD31, phospho‐adenosine 5’‐monophosphate (AMP)‐activated protein kinase (p‐AMPK), AMPK, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) or β‐actin (Cell Signaling Technology). .. After 3 washes with Tris‐buffered saline with Tween‐20 (TBST), the membranes were incubated with the corresponding HRP‐conjugated secondary antibody (Cell Signaling Technology) at room temperature for 1 hour.

    Incubation:

    Article Title: The Cooperative Induction of CCL4 in Human Monocytic Cells by TNF-α and Palmitate Requires MyD88 and Involves MAPK/NF-κB Signaling Pathways
    Article Snippet: Proteins were transferred by electro blotting to an Immuno-Blot PVDF membrane (Bio-Rad Laboratories). .. The membranes were blocked by treating with 5% non-fat milk in PBS for 1 h, followed by overnight incubation at 4 °C with primary antibodies (1:1000 diluted) against p-ERK1/2, p-c-Jun, p-SAPK/JNK, p-NF-κB, ERK1/2, c-Jun, SAPK/JNK and NF-κB (Cell Signaling Technology Inc., MA, USA). .. Blots were washed thrice by using Tris buffered saline with 0.1% TBST and incubated for 2 h with an HRP-conjugated secondary antibody (Promega, WI, USA).

    Article Title: FGF21 promotes ischaemic angiogenesis and endothelial progenitor cells function under diabetic conditions in an AMPK/NAD+‐dependent manner, et al. FGF21 promotes ischaemic angiogenesis and endothelial progenitor cells function under diabetic conditions in an AMPK/NAD+‐dependent manner
    Article Snippet: After determining the protein concentration with Quick Start Bradford Dye Reagent (Bio‐Rad), proteins were separated on a 10% SDS‐PAGE and transferred onto nitrocellulose membranes (Bio‐Rad). .. After blocking in 5% skim milk for 1 hour, the membranes were incubated overnight at 4°C with antibodies against phospho‐extracellular regulated protein kinases 1/2 (p‐Erk1/2), Erk1/2, CD38, NAMPT, poly (ADP‐ribose) polymerase (PARP), CD31, phospho‐adenosine 5’‐monophosphate (AMP)‐activated protein kinase (p‐AMPK), AMPK, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) or β‐actin (Cell Signaling Technology). .. After 3 washes with Tris‐buffered saline with Tween‐20 (TBST), the membranes were incubated with the corresponding HRP‐conjugated secondary antibody (Cell Signaling Technology) at room temperature for 1 hour.

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  • 99
    Cell Signaling Technology Inc erk1 2
    Protein analysis of <t>ERK1/2</t> phosphorylation and FosB expression in striatal tissue. Protein levels were evaluated by western blotting the striatal extracts from each group of mice. (A, B) Phospho-ERK1/2 (A) and FosB (B) signals were observed in the ipsilateral sides of striata. Representative bar graph showing optical densities from the blotting experiments. *P
    Erk1 2, supplied by Cell Signaling Technology Inc, 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/erk1 2/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    erk1 2 - by Bioz Stars, 2021-06
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    97
    Cell Signaling Technology Inc anti erk1 2
    TBL1XR1 enhances GC cell proliferation, migration and invasion via the <t>ERK1/2</t> signalling pathway. ( a ) The expression of TBL1XR1 and pERK1/2 in 134 pairs human GC tissues were assessed by IHC. The representative images show the expression of TBL1XR1 and pERK1/2 in intestinal and diffuse type of GC (scale bar=100 μm). ( b ) Spearman correlation was used to analyse the correlation between TBL1XR1 and pERK1/2 expression in 134 pairs of GC tissues. ( c ) The expression of pERK1/2, ERK1/2 and TBL1XR1 in four GC cell lines were determined by western blot analysis. ( d ) ERK1/2 specific inhibitor U0126 (20 μ M ) suppressed the proliferation of NCI-N87 and SGC7901 cells. ( e ) and ( f ) U0126 (20 μM) inhibited the migratory and invasive ability of NCI-N87 and SGC7901 cells (scale bar=100 μm). ( g ) U0126 (20 μ M ) inhibited the phosphorylation of ERK1/2 and EMT changes in NCI-N87 cells and SGC7901 cells. ( h ) The pERK1/2 level in NCI-N87/TBL1XR1-shRNA and SGC7901/TBL1XR1-shRNA cells was determined by western blot analysis. ( i ) The effect of TBL1XR1 overexpression on pERK1/2 level in BGC823 and MKN45 cells. ( j ) BGC823/TBL1XR1 and MKN45/TBL1XR1 cell proliferation were measured in the presence of U026 (20 μ M ). ( k ) and ( l ) The effect of U0126 on the migratory and invasive ability of BGC823/TBL1XR1 and MKN45/TBL1XR1 cells was analysed and representative images are shown (magnification: × 100, scale bar=100 μm). ( m ) The effect of U0126 (20 μ M ) on the EMT changes of BGC823/TBL1XR1 and MKN45/TBL1XR1 cells was analysed by western blot analysis. Densitometry shows relative protein expression normalized for GAPDH. Data are representative of three independent experiments (mean±s.d.). * P
    Anti Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti erk1 2/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
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    anti erk1 2 - by Bioz Stars, 2021-06
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    N/A
    This peptide is used to block p44 42 MAPK Erk1 2 137F5 Rabbit mAb 4695 reactivity
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    Image Search Results


    Protein analysis of ERK1/2 phosphorylation and FosB expression in striatal tissue. Protein levels were evaluated by western blotting the striatal extracts from each group of mice. (A, B) Phospho-ERK1/2 (A) and FosB (B) signals were observed in the ipsilateral sides of striata. Representative bar graph showing optical densities from the blotting experiments. *P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Gastrodia elata Blume alleviates L-DOPA-induced dyskinesia by normalizing FosB and ERK activation in a 6-OHDA-lesioned Parkinson's disease mouse model

    doi: 10.1186/1472-6882-14-107

    Figure Lengend Snippet: Protein analysis of ERK1/2 phosphorylation and FosB expression in striatal tissue. Protein levels were evaluated by western blotting the striatal extracts from each group of mice. (A, B) Phospho-ERK1/2 (A) and FosB (B) signals were observed in the ipsilateral sides of striata. Representative bar graph showing optical densities from the blotting experiments. *P

    Article Snippet: The phosphorylation ratio of ERK1/2 was increased significantly in the striata of mice in L-DOPA group and was found to be 224% of the control (P < 0.01, compared to 6-OHDA control group).

    Techniques: Expressing, Western Blot, Mouse Assay

    Loss of CDK5 activity results in the attenuation of the Fak/Akt survival pathway. A – C : INS 832/13 cells were transfected for 48 h with scramble (Sc) or CDK5 siRNAs (50 nmol/L). Protein levels of CDK5, Fak, Akt, Bad, ERK1/2, cleaved caspase-3, and phosphorylation levels of Fak Ser732 , Akt Ser473 , ERK1/2, and Bad Ser136 were analyzed by Western blot ( left panels ). β-Actin was used as loading control. The right panels represent the quantification of the Western blot ( n = 4). * P

    Journal: Diabetes

    Article Title: Cyclin-Dependent Kinase 5 Promotes Pancreatic ?-Cell Survival via Fak-Akt Signaling Pathways

    doi: 10.2337/db10-1048

    Figure Lengend Snippet: Loss of CDK5 activity results in the attenuation of the Fak/Akt survival pathway. A – C : INS 832/13 cells were transfected for 48 h with scramble (Sc) or CDK5 siRNAs (50 nmol/L). Protein levels of CDK5, Fak, Akt, Bad, ERK1/2, cleaved caspase-3, and phosphorylation levels of Fak Ser732 , Akt Ser473 , ERK1/2, and Bad Ser136 were analyzed by Western blot ( left panels ). β-Actin was used as loading control. The right panels represent the quantification of the Western blot ( n = 4). * P

    Article Snippet: Membranes were probed overnight at 4°C with CDK5 (DC17), P-FakSer732 , insulin receptor substrate (IRS) 2 (Millipore/Upstate, St. Louis, MO), C/EBP homologous protein (CHOP) (Sc-575, lot #L057; Santa Cruz Biotechnology, Santa Cruz, CA), p35 (#2680), p35/p25 (#2673), P-BadSer136 , Bad, P-AktSer473 , Akt, P-ERK1/2, ERK1/2, Fak, cleaved caspase-3, PARP, β-actin, and glyceraldehyde-3-phosphate dehydrogenase (Cell Signaling Technology, Beverly, MA) antibodies, followed by incubation with horseradish peroxidase-conjugated secondary antibodies (Zymed Laboratories, South San Francisco, CA).

    Techniques: Activity Assay, Transfection, Western Blot

    Roscovitine treatment decreases Fak/Akt survival pathway and induces apoptosis in rat islets. A – C : Isolated WT rat islets were cultured in the absence or presence of roscovitine 10 μmol/L for 48 h. A and B : Protein levels of Fak, IRS2, Akt, Bad, ERK1/2, cleaved caspase-3, and phosphorylation levels of Fak Ser732 , Akt Ser473 , ERK 1/2, and Bad Ser136 were analyzed by Western blot. GAPDH was used as loading control. The graphs represent the quantification of the Western blots. C : Islet apoptosis was assessed by PI staining. The graph represents the quantification of the islet area positive for PI, expressed in % ( n = 7). * P

    Journal: Diabetes

    Article Title: Cyclin-Dependent Kinase 5 Promotes Pancreatic ?-Cell Survival via Fak-Akt Signaling Pathways

    doi: 10.2337/db10-1048

    Figure Lengend Snippet: Roscovitine treatment decreases Fak/Akt survival pathway and induces apoptosis in rat islets. A – C : Isolated WT rat islets were cultured in the absence or presence of roscovitine 10 μmol/L for 48 h. A and B : Protein levels of Fak, IRS2, Akt, Bad, ERK1/2, cleaved caspase-3, and phosphorylation levels of Fak Ser732 , Akt Ser473 , ERK 1/2, and Bad Ser136 were analyzed by Western blot. GAPDH was used as loading control. The graphs represent the quantification of the Western blots. C : Islet apoptosis was assessed by PI staining. The graph represents the quantification of the islet area positive for PI, expressed in % ( n = 7). * P

    Article Snippet: Membranes were probed overnight at 4°C with CDK5 (DC17), P-FakSer732 , insulin receptor substrate (IRS) 2 (Millipore/Upstate, St. Louis, MO), C/EBP homologous protein (CHOP) (Sc-575, lot #L057; Santa Cruz Biotechnology, Santa Cruz, CA), p35 (#2680), p35/p25 (#2673), P-BadSer136 , Bad, P-AktSer473 , Akt, P-ERK1/2, ERK1/2, Fak, cleaved caspase-3, PARP, β-actin, and glyceraldehyde-3-phosphate dehydrogenase (Cell Signaling Technology, Beverly, MA) antibodies, followed by incubation with horseradish peroxidase-conjugated secondary antibodies (Zymed Laboratories, South San Francisco, CA).

    Techniques: Isolation, Cell Culture, Western Blot, Staining

    TBL1XR1 enhances GC cell proliferation, migration and invasion via the ERK1/2 signalling pathway. ( a ) The expression of TBL1XR1 and pERK1/2 in 134 pairs human GC tissues were assessed by IHC. The representative images show the expression of TBL1XR1 and pERK1/2 in intestinal and diffuse type of GC (scale bar=100 μm). ( b ) Spearman correlation was used to analyse the correlation between TBL1XR1 and pERK1/2 expression in 134 pairs of GC tissues. ( c ) The expression of pERK1/2, ERK1/2 and TBL1XR1 in four GC cell lines were determined by western blot analysis. ( d ) ERK1/2 specific inhibitor U0126 (20 μ M ) suppressed the proliferation of NCI-N87 and SGC7901 cells. ( e ) and ( f ) U0126 (20 μM) inhibited the migratory and invasive ability of NCI-N87 and SGC7901 cells (scale bar=100 μm). ( g ) U0126 (20 μ M ) inhibited the phosphorylation of ERK1/2 and EMT changes in NCI-N87 cells and SGC7901 cells. ( h ) The pERK1/2 level in NCI-N87/TBL1XR1-shRNA and SGC7901/TBL1XR1-shRNA cells was determined by western blot analysis. ( i ) The effect of TBL1XR1 overexpression on pERK1/2 level in BGC823 and MKN45 cells. ( j ) BGC823/TBL1XR1 and MKN45/TBL1XR1 cell proliferation were measured in the presence of U026 (20 μ M ). ( k ) and ( l ) The effect of U0126 on the migratory and invasive ability of BGC823/TBL1XR1 and MKN45/TBL1XR1 cells was analysed and representative images are shown (magnification: × 100, scale bar=100 μm). ( m ) The effect of U0126 (20 μ M ) on the EMT changes of BGC823/TBL1XR1 and MKN45/TBL1XR1 cells was analysed by western blot analysis. Densitometry shows relative protein expression normalized for GAPDH. Data are representative of three independent experiments (mean±s.d.). * P

    Journal: Oncogene

    Article Title: Transducin (β)-like 1 X-linked receptor 1 promotes gastric cancer progression via the ERK1/2 pathway

    doi: 10.1038/onc.2016.352

    Figure Lengend Snippet: TBL1XR1 enhances GC cell proliferation, migration and invasion via the ERK1/2 signalling pathway. ( a ) The expression of TBL1XR1 and pERK1/2 in 134 pairs human GC tissues were assessed by IHC. The representative images show the expression of TBL1XR1 and pERK1/2 in intestinal and diffuse type of GC (scale bar=100 μm). ( b ) Spearman correlation was used to analyse the correlation between TBL1XR1 and pERK1/2 expression in 134 pairs of GC tissues. ( c ) The expression of pERK1/2, ERK1/2 and TBL1XR1 in four GC cell lines were determined by western blot analysis. ( d ) ERK1/2 specific inhibitor U0126 (20 μ M ) suppressed the proliferation of NCI-N87 and SGC7901 cells. ( e ) and ( f ) U0126 (20 μM) inhibited the migratory and invasive ability of NCI-N87 and SGC7901 cells (scale bar=100 μm). ( g ) U0126 (20 μ M ) inhibited the phosphorylation of ERK1/2 and EMT changes in NCI-N87 cells and SGC7901 cells. ( h ) The pERK1/2 level in NCI-N87/TBL1XR1-shRNA and SGC7901/TBL1XR1-shRNA cells was determined by western blot analysis. ( i ) The effect of TBL1XR1 overexpression on pERK1/2 level in BGC823 and MKN45 cells. ( j ) BGC823/TBL1XR1 and MKN45/TBL1XR1 cell proliferation were measured in the presence of U026 (20 μ M ). ( k ) and ( l ) The effect of U0126 on the migratory and invasive ability of BGC823/TBL1XR1 and MKN45/TBL1XR1 cells was analysed and representative images are shown (magnification: × 100, scale bar=100 μm). ( m ) The effect of U0126 (20 μ M ) on the EMT changes of BGC823/TBL1XR1 and MKN45/TBL1XR1 cells was analysed by western blot analysis. Densitometry shows relative protein expression normalized for GAPDH. Data are representative of three independent experiments (mean±s.d.). * P

    Article Snippet: Anti-ERK1/2 (no. 9100S), anti-pERK1/2 (no. 9100S), anti-phospho-β-catenin (Ser675) (no. 9567S), anti-pEGFR (Tyr1068) (no. 3777S) and HRP-conjugated secondary (1:5000) antibodies were purchased from Cell Signaling Technology.

    Techniques: Migration, Expressing, Immunohistochemistry, Western Blot, shRNA, Over Expression

    The activation of ERK1/2 induced by TBL1XR1 is mediated via the β-catenin signalling pathway. ( a ) The phosphorylation levels of β-catenin (p-β-catenin) and ERK1/2 in NCI-N87/TBL1XR1-shRNA and SGC7901/TBL1XR1-shRNA cells were detected by western blot analysis. ( b ) The phosphorylation levels of β-catenin and ERK1/2 in BGC823/TBL1XR1 and MKN45/TBL1XR1 cells were detected by western blot analysis. ( c ) The phosphorylation level of ERK1/2 and EMT in BGC823/TBL1XR1 and MKN45/TBL1XR1 cells treated with XAV939 were detected by western blot analysis (10 μ M ). ( d ) The effect of U0126 (20 μ M ) on phosphorylation of β-catenin in NCI-N87 and SGC7901 cells was analysed by western blot analysis. Densitometry shows relative protein expression.

    Journal: Oncogene

    Article Title: Transducin (β)-like 1 X-linked receptor 1 promotes gastric cancer progression via the ERK1/2 pathway

    doi: 10.1038/onc.2016.352

    Figure Lengend Snippet: The activation of ERK1/2 induced by TBL1XR1 is mediated via the β-catenin signalling pathway. ( a ) The phosphorylation levels of β-catenin (p-β-catenin) and ERK1/2 in NCI-N87/TBL1XR1-shRNA and SGC7901/TBL1XR1-shRNA cells were detected by western blot analysis. ( b ) The phosphorylation levels of β-catenin and ERK1/2 in BGC823/TBL1XR1 and MKN45/TBL1XR1 cells were detected by western blot analysis. ( c ) The phosphorylation level of ERK1/2 and EMT in BGC823/TBL1XR1 and MKN45/TBL1XR1 cells treated with XAV939 were detected by western blot analysis (10 μ M ). ( d ) The effect of U0126 (20 μ M ) on phosphorylation of β-catenin in NCI-N87 and SGC7901 cells was analysed by western blot analysis. Densitometry shows relative protein expression.

    Article Snippet: Anti-ERK1/2 (no. 9100S), anti-pERK1/2 (no. 9100S), anti-phospho-β-catenin (Ser675) (no. 9567S), anti-pEGFR (Tyr1068) (no. 3777S) and HRP-conjugated secondary (1:5000) antibodies were purchased from Cell Signaling Technology.

    Techniques: Activation Assay, shRNA, Western Blot, Expressing

    Schematic model of TBL1XR1-promoting ERK activation and gastric cancer progression. GC cell-derived TBL1XR1 promotes the phosphorylation of ERK1/2 via activating β-catenin/MMP7/EGFR, which facilitates the EMT, migration, invasion and metastatic potential of GC cells.

    Journal: Oncogene

    Article Title: Transducin (β)-like 1 X-linked receptor 1 promotes gastric cancer progression via the ERK1/2 pathway

    doi: 10.1038/onc.2016.352

    Figure Lengend Snippet: Schematic model of TBL1XR1-promoting ERK activation and gastric cancer progression. GC cell-derived TBL1XR1 promotes the phosphorylation of ERK1/2 via activating β-catenin/MMP7/EGFR, which facilitates the EMT, migration, invasion and metastatic potential of GC cells.

    Article Snippet: Anti-ERK1/2 (no. 9100S), anti-pERK1/2 (no. 9100S), anti-phospho-β-catenin (Ser675) (no. 9567S), anti-pEGFR (Tyr1068) (no. 3777S) and HRP-conjugated secondary (1:5000) antibodies were purchased from Cell Signaling Technology.

    Techniques: Activation Assay, Derivative Assay, Migration

    The activation of ERK1/2 induced by TBL1XR1 is mediated via the β-catenin/MMP7/EGFR signalling pathway. ( a ) and ( b ) The expression levels of TBL1XR1, p-β-catenin, MMP7, pEGFR and pERK1/2 in NCI-N87/TBL1XR1-shRNA cells ( a ) and BGC823/TBL1XR1 cells ( b ) were detected by western blot analysis. ( c ) and ( d ) The expression levels of TBL1XR1, p-β-catenin, MMP7, pEGFR and pERK1/2 in NCI-N87 cells ( c ) and BGC823/TBL1XR1 cells ( d ) in the presence of XAV939 (10 μ M ), Batimastat (10 μ M ), Afatinib (5 μ M ) and U0126 (20 μ M ) were detected by western blot analysis. Densitometry shows relative protein expression.

    Journal: Oncogene

    Article Title: Transducin (β)-like 1 X-linked receptor 1 promotes gastric cancer progression via the ERK1/2 pathway

    doi: 10.1038/onc.2016.352

    Figure Lengend Snippet: The activation of ERK1/2 induced by TBL1XR1 is mediated via the β-catenin/MMP7/EGFR signalling pathway. ( a ) and ( b ) The expression levels of TBL1XR1, p-β-catenin, MMP7, pEGFR and pERK1/2 in NCI-N87/TBL1XR1-shRNA cells ( a ) and BGC823/TBL1XR1 cells ( b ) were detected by western blot analysis. ( c ) and ( d ) The expression levels of TBL1XR1, p-β-catenin, MMP7, pEGFR and pERK1/2 in NCI-N87 cells ( c ) and BGC823/TBL1XR1 cells ( d ) in the presence of XAV939 (10 μ M ), Batimastat (10 μ M ), Afatinib (5 μ M ) and U0126 (20 μ M ) were detected by western blot analysis. Densitometry shows relative protein expression.

    Article Snippet: Anti-ERK1/2 (no. 9100S), anti-pERK1/2 (no. 9100S), anti-phospho-β-catenin (Ser675) (no. 9567S), anti-pEGFR (Tyr1068) (no. 3777S) and HRP-conjugated secondary (1:5000) antibodies were purchased from Cell Signaling Technology.

    Techniques: Activation Assay, Expressing, shRNA, Western Blot

    TFPI-2 mediates the EGFR-ERK1/2 signaling pathway. ( A ) Left: representative western blots showing the expressions of EGFR, pEGFR, P38, pP38, pMEK1, ERK1/2, and pERK1/2 proteins in cell lysates of MDA231 cells. β-actin was used as a loading control. Right: relative levels of the proteins were normalized to β-actin. Data for statistical analyses was from three independent experiments. * P

    Journal: Scientific Reports

    Article Title: TFPI-2 suppresses breast cancer cell proliferation and invasion through regulation of ERK signaling and interaction with actinin-4 and myosin-9

    doi: 10.1038/s41598-018-32698-3

    Figure Lengend Snippet: TFPI-2 mediates the EGFR-ERK1/2 signaling pathway. ( A ) Left: representative western blots showing the expressions of EGFR, pEGFR, P38, pP38, pMEK1, ERK1/2, and pERK1/2 proteins in cell lysates of MDA231 cells. β-actin was used as a loading control. Right: relative levels of the proteins were normalized to β-actin. Data for statistical analyses was from three independent experiments. * P

    Article Snippet: Anti-EGFR, anti-phospho-EGFR, anti-ERK1/2, anti- phospho-ERK1/2 (pERK1/2), anti-phospho-MEK1/2, anti-P38, and anti- phospho-P38 antibodies were purchased from Cell Signaling, Inc. (MA, USA).

    Techniques: Western Blot