phosphorylated erk1 2  (Cell Signaling Technology Inc)

 
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    Name:
    Phospho p44 42 MAPK Erk1 2 Thr202 Tyr204 197G2 Rabbit 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:
    4377
    Price:
    None
    Category:
    Primary Antibodies
    Source:
    Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase.
    Reactivity:
    Human Mouse Rat Monkey Mink D melanogaster Zebrafish Pig
    Applications:
    Western Blot, Immunofluorescence, Flow Cytometry
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    Structured Review

    Cell Signaling Technology Inc phosphorylated erk1 2
    Inhibiting dynamin eliminates sustained activation of <t>ERK1/2</t> by CXCR7 A) Confocal microscope images of 293T cells transfected transiently with CXCR7-WT-GFP along with dominant negative K44A dynamin (K44A) or control plasmid. Scale bar denotes 20 μm. B) Representative line profile analyses of fluorescence intensity and distribution across cells depicted in panel A. C) Graph depicts bioluminescence from interaction of CXCR7-NLuc with β-arrestin-2-CLuc in transiently transfected 293T cells co-transfected with dominant negative K44A dynamin or control plasmid. Cells were incubated with 100 ng/ml CXCL12 for 30 minutes before quantifying bioluminescence. Data were graphed as mean values + SEM for firefly luciferase photon flux normalized to co-transfected Gaussia luciferase. *, p
    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/phosphorylated erk1 2/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphorylated erk1 2 - by Bioz Stars, 2021-06
    97/100 stars

    Images

    1) Product Images from "Carboxy-terminus of CXCR7 Regulates Receptor Localization and Function"

    Article Title: Carboxy-terminus of CXCR7 Regulates Receptor Localization and Function

    Journal: The international journal of biochemistry & cell biology

    doi: 10.1016/j.biocel.2012.01.007

    Inhibiting dynamin eliminates sustained activation of ERK1/2 by CXCR7 A) Confocal microscope images of 293T cells transfected transiently with CXCR7-WT-GFP along with dominant negative K44A dynamin (K44A) or control plasmid. Scale bar denotes 20 μm. B) Representative line profile analyses of fluorescence intensity and distribution across cells depicted in panel A. C) Graph depicts bioluminescence from interaction of CXCR7-NLuc with β-arrestin-2-CLuc in transiently transfected 293T cells co-transfected with dominant negative K44A dynamin or control plasmid. Cells were incubated with 100 ng/ml CXCL12 for 30 minutes before quantifying bioluminescence. Data were graphed as mean values + SEM for firefly luciferase photon flux normalized to co-transfected Gaussia luciferase. *, p
    Figure Legend Snippet: Inhibiting dynamin eliminates sustained activation of ERK1/2 by CXCR7 A) Confocal microscope images of 293T cells transfected transiently with CXCR7-WT-GFP along with dominant negative K44A dynamin (K44A) or control plasmid. Scale bar denotes 20 μm. B) Representative line profile analyses of fluorescence intensity and distribution across cells depicted in panel A. C) Graph depicts bioluminescence from interaction of CXCR7-NLuc with β-arrestin-2-CLuc in transiently transfected 293T cells co-transfected with dominant negative K44A dynamin or control plasmid. Cells were incubated with 100 ng/ml CXCL12 for 30 minutes before quantifying bioluminescence. Data were graphed as mean values + SEM for firefly luciferase photon flux normalized to co-transfected Gaussia luciferase. *, p

    Techniques Used: Activation Assay, Microscopy, Transfection, Dominant Negative Mutation, Plasmid Preparation, Fluorescence, Incubation, Luciferase

    CXCR7-322 blocks activation of ERK1/2 in response to CXCL12 A) Western blot of 293T cells transfected with CXCR7-GFP or CXCR7-322-GFP following treatment with 100 ng/ml CXCL12 for various periods of time through 30 minutes. Blots show phosphorylated and total ERK1/2, respectively. The gel format minimized separation between bands for ERK1 and ERK2. B) Graph displays relative intensities of phosphorylated to total ERK1/2 for each samples expressed as ratios of arbitrary units.
    Figure Legend Snippet: CXCR7-322 blocks activation of ERK1/2 in response to CXCL12 A) Western blot of 293T cells transfected with CXCR7-GFP or CXCR7-322-GFP following treatment with 100 ng/ml CXCL12 for various periods of time through 30 minutes. Blots show phosphorylated and total ERK1/2, respectively. The gel format minimized separation between bands for ERK1 and ERK2. B) Graph displays relative intensities of phosphorylated to total ERK1/2 for each samples expressed as ratios of arbitrary units.

    Techniques Used: Activation Assay, Western Blot, Transfection

    Related Articles

    Incubation:

    Article Title: Carboxy-terminus of CXCR7 Regulates Receptor Localization and Function
    Article Snippet: .. Lysates were incubated overnight at 4°C with a rabbit monoclonal antibody to phosphorylated ERK1/2 (1:500 dilution; Cell Signaling, antibody 197G2). .. Bound primary antibody was detected with an anti-rabbit secondary antibody linked to horseradish peroxidase (1:5000 dilution; Sigma) incubated for 1 hour at room temperature.

    Article Title: Intrauterine human chorionic gonadotropin infusion in oocyte donors promotes endometrial synchrony and induction of early decidual markers for stromal survival: a randomized clinical trial
    Article Snippet: Each tissue section was dewaxed, rehydrated in graded alcohol series, followed by heat-mediated antigen retrieval in citrate buffer (Antigen unmasking solution, H-3300; Vector Laboratories, Burlingame, CA). .. Slides were blocked for 1 h in 10% normal horse serum (Vector Laboratories; S-2000) in PBS then incubated overnight at 4°C in one of the following primary antibodies: mouse anti-ERα (750 ng/ml; Vector Laboratories; VP-E613), rabbit anti-PR (12 µg/ml; DAKO, Carpinteria, CA; A0098), rabbit anti-ERK1/2 (80 ng/ml; Cell Signaling, Danvers, MA; cs4695), rabbit anti-phospho-ERK1/2 (50 ng/ml; Cell Signaling; cs-4376), rabbit anti-α-smooth muscle actin (233 ng/ml; DAKO; M0851), rabbit anti-NOTCH1 (1 µg/ml; Santa Cruz Biotechnology, Santa Cruz, CA; sc-6014-R) or goat anti-C3 (400 ng/ml; Santa Cruz Biotechnology; sc-14612). .. Subsequently, sections were incubated in respective biotinylated secondary antibodies for anti-mouse, anti-rabbit or anti-goat (Vector Laboratories; BA-9200, BA-1000, BA-9500) followed by HRP-conjugated streptavidin.

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    Cell Signaling Technology Inc antibodies against erk1 2
    Involvement of Bcr-Abl and CK2 in <t>MEK/ERK1/2</t> and PI3K/Akt/mTOR/p70S6K pathways Figure highlights signalling proteins affected by Bcr-Abl and CK2. Arrows and tee-arrows indicate substrate-protein activation or inactivation, respectively. Dashed arrows mean that CK2-phosphorylation affects the protein translocation. Red arrows indicate that, as demonstrated by our results, CK2 promotes rpS6 phosphorylation acting downstream of p70S6K.
    Antibodies Against 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/antibodies against erk1 2/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibodies against erk1 2 - by Bioz Stars, 2021-06
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    94
    Cell Signaling Technology Inc rabbit anti phospho erk1 2 mab
    Increased phosphorylation of p38 MAPK and <t>ERK1/2</t> in RBL-CCR1 cells stimulated via FcϵRI and CCR1. After sensitization with anti-DNP-IgE mAb, RBL-CCR1 cells were stimulated with 10 ng/ml DNP-HSA and/or 100 ng/ml rCCL3 for 5 min. To inhibit p38
    Rabbit Anti Phospho Erk1 2 Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti phospho erk1 2 mab/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho erk1 2 mab - by Bioz Stars, 2021-06
    94/100 stars
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    97
    Cell Signaling Technology Inc anti total erk1 2
    IQGAP1 mediates the formation of a large signalosome in the perinuclear region of the activated NK cells. IL-2–cultured splenic Rap1b +/+ (A) and Rap1b −/− (B) NK cells were activated with anti-NKG2D (A10) for 30 min and stained for IQGAP1 (red), <t>phospho-ERK1/2</t> (green), and DAPI (blue). Three dimensional reconstruction of images was done using 40 individual Z-stacks that were 0.4 µm thick, using FV1-ASW2.0 software. Data shown are one representative panel of 20 images analyzed from three independent experiments.
    Anti Total 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 total erk1 2/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti total erk1 2 - by Bioz Stars, 2021-06
    97/100 stars
      Buy from Supplier

    Image Search Results


    Involvement of Bcr-Abl and CK2 in MEK/ERK1/2 and PI3K/Akt/mTOR/p70S6K pathways Figure highlights signalling proteins affected by Bcr-Abl and CK2. Arrows and tee-arrows indicate substrate-protein activation or inactivation, respectively. Dashed arrows mean that CK2-phosphorylation affects the protein translocation. Red arrows indicate that, as demonstrated by our results, CK2 promotes rpS6 phosphorylation acting downstream of p70S6K.

    Journal: Oncotarget

    Article Title: Inhibition of protein kinase CK2 by CX-5011 counteracts imatinib-resistance preventing rpS6 phosphorylation in chronic myeloid leukaemia cells: new combined therapeutic strategies

    doi: 10.18632/oncotarget.7569

    Figure Lengend Snippet: Involvement of Bcr-Abl and CK2 in MEK/ERK1/2 and PI3K/Akt/mTOR/p70S6K pathways Figure highlights signalling proteins affected by Bcr-Abl and CK2. Arrows and tee-arrows indicate substrate-protein activation or inactivation, respectively. Dashed arrows mean that CK2-phosphorylation affects the protein translocation. Red arrows indicate that, as demonstrated by our results, CK2 promotes rpS6 phosphorylation acting downstream of p70S6K.

    Article Snippet: Antibodies against ERK1/2, phospho-ERK1/2(T202/Y204), phospho-RSK(T573), phospho-Akt(T308), phospho-Akt(S473), phospho-mTOR(S2448), GSK3β, phospho-GSK3β(S9), phospho-rpS6(S235-6), p70S6K, phospho-p70S6K(T389), 4E-BP1 and phospho-4E-BP1(T37/46) were from Cell Signalling Technology (Danvers, MA).

    Techniques: Activation Assay, Translocation Assay

    Analysis of MEK/ERK1/2 and PI3K/Akt/mTOR pathways in imatinib-sensitive and -resistant CML cells A, B, C. S- and R-KCL22, and D. S- and R-K562 cells were lysed and lysate proteins were analysed by western blot with the indicated antibodies. Means of the densitometric values ± SD relative to the phosphorylation extent of ERK1/2 p-T202/Y204 (A, D), Akt p-S473 (B, D) and rpS6 p-S235-236 (C, D) are reported above the immunostained bands. Densitometric values of imatinib-sensitive cells were arbitrarily set equal to 1.0. Figure is representative of at least four separate experiments. *p

    Journal: Oncotarget

    Article Title: Inhibition of protein kinase CK2 by CX-5011 counteracts imatinib-resistance preventing rpS6 phosphorylation in chronic myeloid leukaemia cells: new combined therapeutic strategies

    doi: 10.18632/oncotarget.7569

    Figure Lengend Snippet: Analysis of MEK/ERK1/2 and PI3K/Akt/mTOR pathways in imatinib-sensitive and -resistant CML cells A, B, C. S- and R-KCL22, and D. S- and R-K562 cells were lysed and lysate proteins were analysed by western blot with the indicated antibodies. Means of the densitometric values ± SD relative to the phosphorylation extent of ERK1/2 p-T202/Y204 (A, D), Akt p-S473 (B, D) and rpS6 p-S235-236 (C, D) are reported above the immunostained bands. Densitometric values of imatinib-sensitive cells were arbitrarily set equal to 1.0. Figure is representative of at least four separate experiments. *p

    Article Snippet: Antibodies against ERK1/2, phospho-ERK1/2(T202/Y204), phospho-RSK(T573), phospho-Akt(T308), phospho-Akt(S473), phospho-mTOR(S2448), GSK3β, phospho-GSK3β(S9), phospho-rpS6(S235-6), p70S6K, phospho-p70S6K(T389), 4E-BP1 and phospho-4E-BP1(T37/46) were from Cell Signalling Technology (Danvers, MA).

    Techniques: Western Blot

    Effect of imatinib on MEK/ERK1/2 and PI3K/Akt/mTOR pathways in KCL22 cells S- and R-KCL22 cells, treated with DMSO (Ctrl) or with the indicated amounts of imatinib for 4 h, were lysed and lysate proteins were analysed by western blot with the indicated antibodies. Figure is representative of four separate experiments.

    Journal: Oncotarget

    Article Title: Inhibition of protein kinase CK2 by CX-5011 counteracts imatinib-resistance preventing rpS6 phosphorylation in chronic myeloid leukaemia cells: new combined therapeutic strategies

    doi: 10.18632/oncotarget.7569

    Figure Lengend Snippet: Effect of imatinib on MEK/ERK1/2 and PI3K/Akt/mTOR pathways in KCL22 cells S- and R-KCL22 cells, treated with DMSO (Ctrl) or with the indicated amounts of imatinib for 4 h, were lysed and lysate proteins were analysed by western blot with the indicated antibodies. Figure is representative of four separate experiments.

    Article Snippet: Antibodies against ERK1/2, phospho-ERK1/2(T202/Y204), phospho-RSK(T573), phospho-Akt(T308), phospho-Akt(S473), phospho-mTOR(S2448), GSK3β, phospho-GSK3β(S9), phospho-rpS6(S235-6), p70S6K, phospho-p70S6K(T389), 4E-BP1 and phospho-4E-BP1(T37/46) were from Cell Signalling Technology (Danvers, MA).

    Techniques: Western Blot

    CD147 mediated HCMV-triggered antiviral signaling via the sCyPA-CD147 interaction activated ERK/NF-κB pathway. ( a ) Quantitative RT-PCR analysis of Cyclophilin A (CyPA) mRNA expression kinetics in HFF cells at different time points post-HCMV infection (MOI = 5); ( b ) Western blot analysis of secreted CypA (sCypA) protein levels and intracellular CypA (inCypA) in total protein lysates in culture mediums of HFF cells at the indicated times post HCMV infection (MOI = 5); ( c ) Western blot analysis of the phosphorylation levels of ERK1/2 (p-ERK1/2), NF-κB p65 subunit (p-p65) and IRF3 (p-IRF3) in response to HCMV infection for 6 h (MOI = 0.5) compared to mock infection (Mock) with the medium only. The intensities of bands were quantified by Image Lab 4.0 (Bio-Rad) and relative values normalized to GAPDH are indicated by the numbers under the lanes; ( d ) Analysis of the phosphorylation activation of ERK1/2, induced by HCMV infection for 6 h (MOI = 0.5) with the culture medium, pre-supplemented with cyclosporine (CsA) (5 μg/mL) or CD147 mAb (50 μg/mL).

    Journal: Viruses

    Article Title: Human Cytomegalovirus Encoded miR-US25-1-5p Attenuates CD147/EMMPRIN-Mediated Early Antiviral Response

    doi: 10.3390/v9120365

    Figure Lengend Snippet: CD147 mediated HCMV-triggered antiviral signaling via the sCyPA-CD147 interaction activated ERK/NF-κB pathway. ( a ) Quantitative RT-PCR analysis of Cyclophilin A (CyPA) mRNA expression kinetics in HFF cells at different time points post-HCMV infection (MOI = 5); ( b ) Western blot analysis of secreted CypA (sCypA) protein levels and intracellular CypA (inCypA) in total protein lysates in culture mediums of HFF cells at the indicated times post HCMV infection (MOI = 5); ( c ) Western blot analysis of the phosphorylation levels of ERK1/2 (p-ERK1/2), NF-κB p65 subunit (p-p65) and IRF3 (p-IRF3) in response to HCMV infection for 6 h (MOI = 0.5) compared to mock infection (Mock) with the medium only. The intensities of bands were quantified by Image Lab 4.0 (Bio-Rad) and relative values normalized to GAPDH are indicated by the numbers under the lanes; ( d ) Analysis of the phosphorylation activation of ERK1/2, induced by HCMV infection for 6 h (MOI = 0.5) with the culture medium, pre-supplemented with cyclosporine (CsA) (5 μg/mL) or CD147 mAb (50 μg/mL).

    Article Snippet: We also used anti-HCMV IE1/2 mouse mAb (ab53495, Abcam, Cambridge, UK), rabbit anti-human CyPA mAb (ab3563, Abcam), phospho-MEK1/2 (Ser217/221) rabbit mAb (#9154, Cell Signaling Technology (CST), Danvers, MA, USA), Erk1/2 rabbit mAb (#4695, CST), IRF-3 rabbit mAb (#11904, CST), phospho-IRF-3 (Ser396) rabbit mAb (#29047, CST), NF-κB p65 rabbit mAb (#8242, CST), phospho-NF-κB p65 (Ser536) rabbit mAb (#3033, CST), and anti-GAPDH mouse mAb (60004-1-Ig, Proteintech, Rosemont, IL, USA).

    Techniques: Quantitative RT-PCR, Expressing, Infection, Western Blot, Activation Assay

    General model for HCMV miR-US25-1-5p in attenuating CD147/EMMPRIN-mediated HCMV-triggered early antiviral response. Upon HCMV infection, intracellular CypA is released. As a paracrine proinflammatory cytokine, CyPA interacts with CD147 of those uninfected cells and tissues, and stimulates the phosphorylation of ERK1/2. The activation of NF-κB then occurs, followed by a subsequent activation of the IRF3 phosphorylation and the orchestrated NF-κB-dependent, IRF3 plus NF-κB-dependent immune responses. HCMV-encoded miR-US25-1-5p targeting CD147, CD147-blocking mAb, and an inhibitor of CyPA (CsA) can specifically inhibit this process.

    Journal: Viruses

    Article Title: Human Cytomegalovirus Encoded miR-US25-1-5p Attenuates CD147/EMMPRIN-Mediated Early Antiviral Response

    doi: 10.3390/v9120365

    Figure Lengend Snippet: General model for HCMV miR-US25-1-5p in attenuating CD147/EMMPRIN-mediated HCMV-triggered early antiviral response. Upon HCMV infection, intracellular CypA is released. As a paracrine proinflammatory cytokine, CyPA interacts with CD147 of those uninfected cells and tissues, and stimulates the phosphorylation of ERK1/2. The activation of NF-κB then occurs, followed by a subsequent activation of the IRF3 phosphorylation and the orchestrated NF-κB-dependent, IRF3 plus NF-κB-dependent immune responses. HCMV-encoded miR-US25-1-5p targeting CD147, CD147-blocking mAb, and an inhibitor of CyPA (CsA) can specifically inhibit this process.

    Article Snippet: We also used anti-HCMV IE1/2 mouse mAb (ab53495, Abcam, Cambridge, UK), rabbit anti-human CyPA mAb (ab3563, Abcam), phospho-MEK1/2 (Ser217/221) rabbit mAb (#9154, Cell Signaling Technology (CST), Danvers, MA, USA), Erk1/2 rabbit mAb (#4695, CST), IRF-3 rabbit mAb (#11904, CST), phospho-IRF-3 (Ser396) rabbit mAb (#29047, CST), NF-κB p65 rabbit mAb (#8242, CST), phospho-NF-κB p65 (Ser536) rabbit mAb (#3033, CST), and anti-GAPDH mouse mAb (60004-1-Ig, Proteintech, Rosemont, IL, USA).

    Techniques: Infection, Activation Assay, Blocking Assay

    Increased phosphorylation of p38 MAPK and ERK1/2 in RBL-CCR1 cells stimulated via FcϵRI and CCR1. After sensitization with anti-DNP-IgE mAb, RBL-CCR1 cells were stimulated with 10 ng/ml DNP-HSA and/or 100 ng/ml rCCL3 for 5 min. To inhibit p38

    Journal: The Journal of Biological Chemistry

    Article Title: Evidence That Formation of Vimentin?Mitogen-activated Protein Kinase (MAPK) Complex Mediates Mast Cell Activation following Fc?RI/CC Chemokine Receptor 1 Cross-talk *

    doi: 10.1074/jbc.M111.319624

    Figure Lengend Snippet: Increased phosphorylation of p38 MAPK and ERK1/2 in RBL-CCR1 cells stimulated via FcϵRI and CCR1. After sensitization with anti-DNP-IgE mAb, RBL-CCR1 cells were stimulated with 10 ng/ml DNP-HSA and/or 100 ng/ml rCCL3 for 5 min. To inhibit p38

    Article Snippet: Mouse anti-vimentin phospho-Ser-55 mAb (4A4) , rabbit anti-vimentin phospho-Ser-71 mAb (TM71) , mouse anti-vimentin phospho-Ser-6 (MO6) or vimentin phospho-Ser-82 mAb (MO82) , rabbit anti-vimentin mAb (H54), rabbit anti-phospho-ERK1/2 mAb (Cell Signaling Technology), and rabbit anti-phospho-p38 MAPK mAb (Cell Signaling Technology) were used as the primary antibodies.

    Techniques:

    Hypothetical model for FcϵRI and CCR1 mediated signaling pathways in mast cells. FcϵRI and CCR1 activation synergistically induces phosphorylation of ERK1/2, p38 MAPK and vimentin in mast cells. Phosphorylation of vimentin induces disassembly

    Journal: The Journal of Biological Chemistry

    Article Title: Evidence That Formation of Vimentin?Mitogen-activated Protein Kinase (MAPK) Complex Mediates Mast Cell Activation following Fc?RI/CC Chemokine Receptor 1 Cross-talk *

    doi: 10.1074/jbc.M111.319624

    Figure Lengend Snippet: Hypothetical model for FcϵRI and CCR1 mediated signaling pathways in mast cells. FcϵRI and CCR1 activation synergistically induces phosphorylation of ERK1/2, p38 MAPK and vimentin in mast cells. Phosphorylation of vimentin induces disassembly

    Article Snippet: Mouse anti-vimentin phospho-Ser-55 mAb (4A4) , rabbit anti-vimentin phospho-Ser-71 mAb (TM71) , mouse anti-vimentin phospho-Ser-6 (MO6) or vimentin phospho-Ser-82 mAb (MO82) , rabbit anti-vimentin mAb (H54), rabbit anti-phospho-ERK1/2 mAb (Cell Signaling Technology), and rabbit anti-phospho-p38 MAPK mAb (Cell Signaling Technology) were used as the primary antibodies.

    Techniques: Activation Assay

    IQGAP1 mediates the formation of a large signalosome in the perinuclear region of the activated NK cells. IL-2–cultured splenic Rap1b +/+ (A) and Rap1b −/− (B) NK cells were activated with anti-NKG2D (A10) for 30 min and stained for IQGAP1 (red), phospho-ERK1/2 (green), and DAPI (blue). Three dimensional reconstruction of images was done using 40 individual Z-stacks that were 0.4 µm thick, using FV1-ASW2.0 software. Data shown are one representative panel of 20 images analyzed from three independent experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: Rap1b facilitates NK cell functions via IQGAP1-mediated signalosomes

    doi: 10.1084/jem.20100040

    Figure Lengend Snippet: IQGAP1 mediates the formation of a large signalosome in the perinuclear region of the activated NK cells. IL-2–cultured splenic Rap1b +/+ (A) and Rap1b −/− (B) NK cells were activated with anti-NKG2D (A10) for 30 min and stained for IQGAP1 (red), phospho-ERK1/2 (green), and DAPI (blue). Three dimensional reconstruction of images was done using 40 individual Z-stacks that were 0.4 µm thick, using FV1-ASW2.0 software. Data shown are one representative panel of 20 images analyzed from three independent experiments.

    Article Snippet: The blots were probed with following antibodies: rabbit anti–total Rap1 (Santa Cruz Biotechnology, Inc.), anti-Rap1a (Cell Signaling Technology), rabbit anti-Rap1b (Cell Signaling Technology), anti-GST mAb (Santa Cruz Biotechnology, Inc.), anti–phospho-Vav-1 (Y160; Abcam), anti–phospho-PAK1/2/3 (S141; Abcam), anti–phospho-p38 (Thr180/Tyr182; Cell Signaling Technology), anti–phospho-ERK1/2 (Thr202/Tyr204; Cell Signaling Technology), anti-phospho-JNK1/2 (Thr183/Tyr185; Cell Signaling Technology), anti–total Vav-1 (Cell Signaling Technology), anti–total PAK1/2/3 (Millipore), anti–total p38 (Cell Signaling Technology), anti–total ERK1/2 (Cell Signaling Technology), anti-total JNK1/2 (Cell Signaling Technology), anti-IQGAP1 (BD), and anti–β-Actin (Millipore) mAbs.

    Techniques: Cell Culture, Staining, Software

    Rap1b regulates IQGAP1-mediated signaling cascade. (A) IL-2–cultured splenic NK cells were activated with anti-NKG2D for 30 min and stained for total Rap1 (green), IQGAP1 (red), and DAPI (blue) and analyzed by confocal microscopy. Inserts in the bottom panels are the enlarged views of the select NK cells. (B and C) NK cells from Rap1b +/+ or Rap1b −/− mice were stimulated with anti-NKG2D for the indicated periods and the phosphorylation of Vav-1 (B) and Pak-1/2/3 (C) was analyzed. (D) IL-2–cultured splenic NK cells from Rap1b +/+ mice were stimulated with anti-NKG2D (A10; 5 µg/ml). Cell lysates were subjected to immunoprecipitation (IP) with anti–B-Raf or pull down (PD) using recombinant GST-C-Raf. Resulting IP or PD were probed for the presence of Rap1-GTP or Rap1b-GTP. One representative experiment out of three is shown. (E and F) IL-2–cultured NK cells were stimulated with anti-NKG2D, and the cell lysates were analyzed for phospho and total proteins of B-Raf and C-Raf (E) or p38, JNK1/2, and ERK1/2 (F). Data presented in A–F were one representative of three independent experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: Rap1b facilitates NK cell functions via IQGAP1-mediated signalosomes

    doi: 10.1084/jem.20100040

    Figure Lengend Snippet: Rap1b regulates IQGAP1-mediated signaling cascade. (A) IL-2–cultured splenic NK cells were activated with anti-NKG2D for 30 min and stained for total Rap1 (green), IQGAP1 (red), and DAPI (blue) and analyzed by confocal microscopy. Inserts in the bottom panels are the enlarged views of the select NK cells. (B and C) NK cells from Rap1b +/+ or Rap1b −/− mice were stimulated with anti-NKG2D for the indicated periods and the phosphorylation of Vav-1 (B) and Pak-1/2/3 (C) was analyzed. (D) IL-2–cultured splenic NK cells from Rap1b +/+ mice were stimulated with anti-NKG2D (A10; 5 µg/ml). Cell lysates were subjected to immunoprecipitation (IP) with anti–B-Raf or pull down (PD) using recombinant GST-C-Raf. Resulting IP or PD were probed for the presence of Rap1-GTP or Rap1b-GTP. One representative experiment out of three is shown. (E and F) IL-2–cultured NK cells were stimulated with anti-NKG2D, and the cell lysates were analyzed for phospho and total proteins of B-Raf and C-Raf (E) or p38, JNK1/2, and ERK1/2 (F). Data presented in A–F were one representative of three independent experiments.

    Article Snippet: The blots were probed with following antibodies: rabbit anti–total Rap1 (Santa Cruz Biotechnology, Inc.), anti-Rap1a (Cell Signaling Technology), rabbit anti-Rap1b (Cell Signaling Technology), anti-GST mAb (Santa Cruz Biotechnology, Inc.), anti–phospho-Vav-1 (Y160; Abcam), anti–phospho-PAK1/2/3 (S141; Abcam), anti–phospho-p38 (Thr180/Tyr182; Cell Signaling Technology), anti–phospho-ERK1/2 (Thr202/Tyr204; Cell Signaling Technology), anti-phospho-JNK1/2 (Thr183/Tyr185; Cell Signaling Technology), anti–total Vav-1 (Cell Signaling Technology), anti–total PAK1/2/3 (Millipore), anti–total p38 (Cell Signaling Technology), anti–total ERK1/2 (Cell Signaling Technology), anti-total JNK1/2 (Cell Signaling Technology), anti-IQGAP1 (BD), and anti–β-Actin (Millipore) mAbs.

    Techniques: Cell Culture, Staining, Confocal Microscopy, Mouse Assay, Immunoprecipitation, Recombinant

    Rap1b regulates IQGAP1-mediated signalosome formation and ERK1/2 phosphorylation. (A) anti-NKG2D or (B) unstimulated splenic NK cells were stained for phospho ERK1/2 (green), IQGAP1 (red), and DAPI (blue), and analyzed by confocal microscopy. Data presented were one representative set from of > 3 independent experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: Rap1b facilitates NK cell functions via IQGAP1-mediated signalosomes

    doi: 10.1084/jem.20100040

    Figure Lengend Snippet: Rap1b regulates IQGAP1-mediated signalosome formation and ERK1/2 phosphorylation. (A) anti-NKG2D or (B) unstimulated splenic NK cells were stained for phospho ERK1/2 (green), IQGAP1 (red), and DAPI (blue), and analyzed by confocal microscopy. Data presented were one representative set from of > 3 independent experiments.

    Article Snippet: The blots were probed with following antibodies: rabbit anti–total Rap1 (Santa Cruz Biotechnology, Inc.), anti-Rap1a (Cell Signaling Technology), rabbit anti-Rap1b (Cell Signaling Technology), anti-GST mAb (Santa Cruz Biotechnology, Inc.), anti–phospho-Vav-1 (Y160; Abcam), anti–phospho-PAK1/2/3 (S141; Abcam), anti–phospho-p38 (Thr180/Tyr182; Cell Signaling Technology), anti–phospho-ERK1/2 (Thr202/Tyr204; Cell Signaling Technology), anti-phospho-JNK1/2 (Thr183/Tyr185; Cell Signaling Technology), anti–total Vav-1 (Cell Signaling Technology), anti–total PAK1/2/3 (Millipore), anti–total p38 (Cell Signaling Technology), anti–total ERK1/2 (Cell Signaling Technology), anti-total JNK1/2 (Cell Signaling Technology), anti-IQGAP1 (BD), and anti–β-Actin (Millipore) mAbs.

    Techniques: Staining, Confocal Microscopy