sapk jnk antibody  (Cell Signaling Technology Inc)

 
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    Name:
    SAPK JNK Antibody
    Description:
    The stress activated protein kinase Jun amino terminal kinase SAPK JNK is potently and preferentially activated by a variety of environmental stresses including UV and gamma radiation ceramides inflammatory cytokines and in some instances growth factors and GPCR agonists 1 6 As with the other MAPKs the core signaling unit is composed of a MAPKKK typically MEKK1 MEKK4 or by one of the mixed lineage kinases MLKs which phosphorylate and activate MKK4 7 Upon activation MKKs phosphorylate and activate the SAPK JNK kinase 2 Stress signals are delivered to this cascade by small GTPases of the Rho family Rac Rho cdc42 3 Both Rac1 and cdc42 mediate the stimulation of MEKKs and MLKs 3 Alternatively MKK4 7 can be activated in a GTPase independent mechanism via stimulation of a germinal center kinase GCK family member 4 There are three SAPK JNK genes each of which undergoes alternative splicing resulting in numerous isoforms 3 SAPK JNK when active as a dimer can translocate to the nucleus and regulate transcription through its effects on c Jun ATF 2 and other transcription factors 3 5
    Catalog Number:
    9252
    Price:
    None
    Applications:
    Western Blot
    Category:
    Primary Antibodies
    Source:
    Polyclonal antibodies are produced by immunizing animals with a recombinant human JNK2 fusion protein. Antibodies are purified by protein A and peptide affinity chromatography.
    Reactivity:
    Human Mouse Rat Hamster Monkey Zebrafish Bovine S cerevisiae
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    Structured Review

    Cell Signaling Technology Inc sapk jnk antibody
    NK-4 attenuated H 2 O 2 -induced <t>SAPK/JNK</t> phosphorylation in PC12 cells. PC12 cells were treated with 400 µM H 2 O 2 in the presence of the indicated concentrations of NK-4 for 2 hr. Whole cell lysates were analyzed by Western blotting using anti-phospho-SAPK/JNK antibody (upper panel), or anti-SAPK/JNK antibody (bottom panel). The graph shows the ratio of phosphorylated SAPK/JNK to total SAPK/JNK at each time point.
    The stress activated protein kinase Jun amino terminal kinase SAPK JNK is potently and preferentially activated by a variety of environmental stresses including UV and gamma radiation ceramides inflammatory cytokines and in some instances growth factors and GPCR agonists 1 6 As with the other MAPKs the core signaling unit is composed of a MAPKKK typically MEKK1 MEKK4 or by one of the mixed lineage kinases MLKs which phosphorylate and activate MKK4 7 Upon activation MKKs phosphorylate and activate the SAPK JNK kinase 2 Stress signals are delivered to this cascade by small GTPases of the Rho family Rac Rho cdc42 3 Both Rac1 and cdc42 mediate the stimulation of MEKKs and MLKs 3 Alternatively MKK4 7 can be activated in a GTPase independent mechanism via stimulation of a germinal center kinase GCK family member 4 There are three SAPK JNK genes each of which undergoes alternative splicing resulting in numerous isoforms 3 SAPK JNK when active as a dimer can translocate to the nucleus and regulate transcription through its effects on c Jun ATF 2 and other transcription factors 3 5
    https://www.bioz.com/result/sapk jnk antibody/product/Cell Signaling Technology Inc
    Average 99 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    sapk jnk antibody - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Neurotrophic Effects of a Cyanine Dye via the PI3K-Akt Pathway: Attenuation of Motor Discoordination and Neurodegeneration in an Ataxic Animal Model"

    Article Title: Neurotrophic Effects of a Cyanine Dye via the PI3K-Akt Pathway: Attenuation of Motor Discoordination and Neurodegeneration in an Ataxic Animal Model

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0017137

    NK-4 attenuated H 2 O 2 -induced SAPK/JNK phosphorylation in PC12 cells. PC12 cells were treated with 400 µM H 2 O 2 in the presence of the indicated concentrations of NK-4 for 2 hr. Whole cell lysates were analyzed by Western blotting using anti-phospho-SAPK/JNK antibody (upper panel), or anti-SAPK/JNK antibody (bottom panel). The graph shows the ratio of phosphorylated SAPK/JNK to total SAPK/JNK at each time point.
    Figure Legend Snippet: NK-4 attenuated H 2 O 2 -induced SAPK/JNK phosphorylation in PC12 cells. PC12 cells were treated with 400 µM H 2 O 2 in the presence of the indicated concentrations of NK-4 for 2 hr. Whole cell lysates were analyzed by Western blotting using anti-phospho-SAPK/JNK antibody (upper panel), or anti-SAPK/JNK antibody (bottom panel). The graph shows the ratio of phosphorylated SAPK/JNK to total SAPK/JNK at each time point.

    Techniques Used: Western Blot

    2) Product Images from "Differential Dependence of Regulatory Volume Decrease Behavior in Rabbit Corneal Epithelial Cells on MAPK Superfamily Activation"

    Article Title: Differential Dependence of Regulatory Volume Decrease Behavior in Rabbit Corneal Epithelial Cells on MAPK Superfamily Activation

    Journal:

    doi: 10.1016/j.exer.2007.02.004

    Activation of SAPK/JNK unaffected by Cl − and K + channel inhibition. Activation of SAPK/JNK was probed either in isotonic (control) solution (300 mOsm), or in 150 mOsm hypotonic solution with or without (A) Cl--free solution, NPPB (100 μM),
    Figure Legend Snippet: Activation of SAPK/JNK unaffected by Cl − and K + channel inhibition. Activation of SAPK/JNK was probed either in isotonic (control) solution (300 mOsm), or in 150 mOsm hypotonic solution with or without (A) Cl--free solution, NPPB (100 μM),

    Techniques Used: Activation Assay, Inhibition

    Inhibition of RVD by SAPK/JNK inhibition. RVD was evaluated for 30 min in the absence (control) or presence of 30 μM SP600125 (SP) in response to 150 mOsm hypotonic challenge (underline) (A). The inset indicated SP600125 that inhibited SAPK/JNK.
    Figure Legend Snippet: Inhibition of RVD by SAPK/JNK inhibition. RVD was evaluated for 30 min in the absence (control) or presence of 30 μM SP600125 (SP) in response to 150 mOsm hypotonic challenge (underline) (A). The inset indicated SP600125 that inhibited SAPK/JNK.

    Techniques Used: Inhibition

    Time-dependent activation of Erk1/2, p38 and SAPK/JNK induced by hypotonic challenge. RCEC were exposed to either isotonic (control) solution (300 mOsm) or 150 mOsm hypotonic challenge for the time periods indicated. Activation of Erk1/2 (A), p38 (B),
    Figure Legend Snippet: Time-dependent activation of Erk1/2, p38 and SAPK/JNK induced by hypotonic challenge. RCEC were exposed to either isotonic (control) solution (300 mOsm) or 150 mOsm hypotonic challenge for the time periods indicated. Activation of Erk1/2 (A), p38 (B),

    Techniques Used: Activation Assay

    Osmolarity-dependent activation of Erk1/2, p38 and SAPK/JNK by hypotonic challenges. RCEC were exposed to either an isotonic control solution (300 mOsm), 225, or 150 mOsm hypotonic challenge for 2.5 min for Erk1/2 and SAPK/JNK experiments, or 30 min for
    Figure Legend Snippet: Osmolarity-dependent activation of Erk1/2, p38 and SAPK/JNK by hypotonic challenges. RCEC were exposed to either an isotonic control solution (300 mOsm), 225, or 150 mOsm hypotonic challenge for 2.5 min for Erk1/2 and SAPK/JNK experiments, or 30 min for

    Techniques Used: Activation Assay

    3) Product Images from "Tetra- and Penta-Acylated Lipid A Structures of Porphyromonas gingivalis LPS Differentially Activate TLR4-Mediated NF-?B Signal Transduction Cascade and Immuno-Inflammatory Response in Human Gingival Fibroblasts"

    Article Title: Tetra- and Penta-Acylated Lipid A Structures of Porphyromonas gingivalis LPS Differentially Activate TLR4-Mediated NF-?B Signal Transduction Cascade and Immuno-Inflammatory Response in Human Gingival Fibroblasts

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0058496

    P. gingivalis (Pg) LPS (PgLPS) and E. coli LPS activated the MAPK pathway in HGFs. Kinetics of P38 mitogen activated protein kinase (P38 MAPK), extracellular signal-regulated kinase1/2(ERK1/2), and Stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK) phosphorylation in HGFs are shown in 5.1 , 5.2 and 5.3 , respectively. Cells were treated with PgLPS 1435/1449 (A), PgLPS 1690 (B) and E. coli LPS (C) at 1 µg/mL for the indicated period of time. Cell extracts were prepared and the levels of P38 MAPK, phospho-p38MAPK, ERK, phospho-ERK, JNK and phospho-JNK were determined by western blotting. Quantification of band intensities was performed by densitometry analysis using Image J software. The fold increase values of phospho-protiens of P38 MAPK (5.1D), ERK1/2 (5.2D) and SAPK/JNK (5.3D) as compared with the total protein are shown in the graphs (arbitrary units over control after normalization to the total protein). Equal loading was confirmed by stripping the immunoblot and re-probing it for α-Tubulin. The data shown here are from a representative experiment repeated three times with similar results . *Significant difference with a p -value
    Figure Legend Snippet: P. gingivalis (Pg) LPS (PgLPS) and E. coli LPS activated the MAPK pathway in HGFs. Kinetics of P38 mitogen activated protein kinase (P38 MAPK), extracellular signal-regulated kinase1/2(ERK1/2), and Stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK) phosphorylation in HGFs are shown in 5.1 , 5.2 and 5.3 , respectively. Cells were treated with PgLPS 1435/1449 (A), PgLPS 1690 (B) and E. coli LPS (C) at 1 µg/mL for the indicated period of time. Cell extracts were prepared and the levels of P38 MAPK, phospho-p38MAPK, ERK, phospho-ERK, JNK and phospho-JNK were determined by western blotting. Quantification of band intensities was performed by densitometry analysis using Image J software. The fold increase values of phospho-protiens of P38 MAPK (5.1D), ERK1/2 (5.2D) and SAPK/JNK (5.3D) as compared with the total protein are shown in the graphs (arbitrary units over control after normalization to the total protein). Equal loading was confirmed by stripping the immunoblot and re-probing it for α-Tubulin. The data shown here are from a representative experiment repeated three times with similar results . *Significant difference with a p -value

    Techniques Used: Western Blot, Software, Stripping Membranes

    Related Articles

    Sandwich ELISA:

    Article Title: Mitogen-activated protein kinase pathway mediates N-(4-hydroxyphenyl)-retinamide-induced neuronal differentiation in the ARPE-19 human retinal pigment epithelial cell line
    Article Snippet: .. Rabbit antibodies and phospho-specific antibodies of c-Raf, MEK1/2, ERK1/2, SAPK/JNK, c-Jun, PathScan® Phospho-p44/42 MAPK sandwich ELISA kit, and human specific SignalSilence® p44/p42 MAPK siRNA kit were from Cell Signaling Technologies (Beverly, MA). ..

    Binding Assay:

    Article Title: Cell Stress Induces Upregulation of Osteopontin via the ERK Pathway in Type II Alveolar Epithelial Cells
    Article Snippet: .. Anti-phospho-specific p44/42 mitogen-activated protein kinase (MAPK) (extracellular signal-regulated protein kinase1/2 (ERK1/2)), anti-ERK1/2, anti-phospho-specific stress-activated protein kinase (SAPK)/c-Jun NH2 -terminal kinase (JNK), anti-SAPK/JNK, anti-phospho-specific p38 MAPK, anti-p38 MAPK, anti-β-actin, anti-immunoglobulin heavy-chain binding protein (BiP) antibodies and horseradish peroxidase (HRP)-conjugated secondary antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). .. Recombinant human OPN was purchased from R & D Systems (Minneapolis, MN, USA).

    other:

    Article Title: HCV induces transforming growth factor β1 through activation of endoplasmic reticulum stress and the unfolded protein response
    Article Snippet: Antibodies Each of the primary antibodies were used according to the manufacturer’s protocol: Phospho-JNK #4671, JNK #9252, IRE1α #3294, PERK #5683, phospho-p38 MAPK #9215, p38 MAPK #9212, phospho-p44/42 MAPK #9106, p44/42 MAPK #4695, (Cell Signaling, Danvers, MA). phospho- IRE1α #PA1-16927, phospho-PERK #MA5-15033, and HCV core (Thermo Scientific, Rockford, IL).

    Article Title: Indirubin 3′-Oxime Inhibits Migration, Invasion, and Metastasis InVivo in Mice Bearing Spontaneously Occurring Pancreatic Cancer via Blocking the RAF/ERK, AKT, and SAPK/JNK Pathways
    Article Snippet: PDAC cell lines (#146, #147, and #244) were treated with Indox for 24 h. Antibodies to MMP-2, MMP-9 (1:100; Kyowa Pharma Chemical Co.), B-RAF (1:1000; Abcam); p-B-RAF (Ser446), p-ERK (Tyr204), p-AKT (Thr308), SAPK/JNK, p-SAPK/JNK (Tyr183), and p-c-Jun (Thr91) (1:1000; Cell Signaling Technology); Akt, c-Jun, GAPDH (1:1000; R & D systems); MMP-7 and ERK (1:1000; Santa Cruz) were used.

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  • 85
    Cell Signaling Technology Inc anti phospho sapk jnk igg
    Ischemia/reperfusion-mediated activation of MAPKs in the lung. PKC β +/+ and PKC β –/– mice underwent the indicated period of left-lung I/R. Animals were sacrificed and protein extracts from the I/R and uninstrumented lung were prepared and subjected to SDS-PAGE (12%, 50 ∝g of protein/lane). Immunoblotting with phospho-p44/42 MAPK antibody or total p44/42 MAPK antibody ( B ), phospho-p38 MAPK antibody, or total p38 MAPK antibody ( I ), and <t>phospho-SAPK/JNK</t> antibody or total SAPK/JNK antibody ( J ) was performed. Data are shown as mean ± SEM of five experiments. Immunohistochemical analysis of phospho-p44/42 expression in murine lung from uninstrumented ( B ) or I/R ( C ) PKC β +/+ mice, and from uninstrumented ( D ) or I/R ( E ) PKC β –/– mice was performed. Scale bar: 50 ∝m. I/R lungs from PKC β +/+ mice were subjected to immunofluorescence microscopy and double stained with an anti–phospho-p44/42 antibody (red) ( F ) and an anti-macrophage antibody (F4/80, green) ( G ). The merging of F (pERK1/2) and G (F4/80) is shown in H . Arrows in F and arrowheads in G indicate dually stained cells Original magnification in F – H , ∞1,000. * P
    Anti Phospho Sapk Jnk Igg, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho sapk jnk igg/product/Cell Signaling Technology Inc
    Average 85 stars, based on 1 article reviews
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    91
    Cell Signaling Technology Inc total jnk1 2 sapks
    CFM-4.16 stimulates apoptosis in parental and TKI-resistant NSCLC cells in part by upregulating pro-apoptotic CARP-1 and activating <t>SAPKs</t> ( A – C ) Indicated parental and TKI-resistant NSCLC cells were either untreated (Control), treated with TKI, CFM-4, or CFM-4.16 for noted dose and time. Cell lysates were analyzed by Western blotting (WB) as in Methods for levels of CARP-1, cyclin B1, cleaved PARP and caspase-8, and activation (phosphorylation) of pro-apoptotic p38 and <t>JNK1/2</t> SAPKs. The western blot membranes were subsequently probed with anti-actin or α-tubulin antibodies to assess equal loading. The presence of respective protein is indicated by an arrowhead on the left side of each blot. Approximate location of various molecular weight markers is indicated on the right side of each blot. kDa, kilodalton.
    Total Jnk1 2 Sapks, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/total jnk1 2 sapks/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc anti mouse stress activated protein kinase jnk
    Analysis of signaling pathways involved in rLsa21-mediated cytokine production in mouse macrophages. ( A ) RAW264.7 cells were pretreated for 30 min at 37 °C with anti-TLR2 (30 μg/ml), anti-TLR4 (30 μg/ml), or an isotype control Ab (30 μg/ml) before stimulation with rLsa21 (5 μg/ml) for 30 min. Levels of phosphorylated <t>p38,</t> <t>JNK,</t> and ERK1/2 induced by rLsa21 and degraded IκBα were analyzed by western blot using anti–phospho-p38 (p-p38), or anti–phospho-ERK1/2 (p-ERK1/2), anti–phospho-JNK (p-JNK) and anti-mouse IκBα as well as a specific control Ab for each of the unphosphorylated kinases. Data are representative of those obtained in three independent experiments. ( B ) TLR2 −/− , TLR4 −/− or TLR2 −/− /TLR4 −/− DKO macrophages cell lines were stimulated with rLsa21 (5 μg/ml) for 30 min. Levels of phosphorylated p38, JNK, and ERK1/2 induced by rLsa21 were analyzed by western blot as described in materials and methods. ( C ) RAW 264.7 cells were pretreated for 30 min with NF-kB inhibitor (SN50; 20 μM), JNK inhibitor (SP600125; 40 μM) or p38MAPK inhibitor (SB203580; 30 μM) and then stimulated with rLsa21 (2 μg/ml). After incubation, supernatants were harvested and levels of IL-6 and TNF-α were measured by ELISA. Data are representative of three different experiments. (*Indicates P
    Anti Mouse Stress Activated Protein Kinase Jnk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mouse stress activated protein kinase jnk/product/Cell Signaling Technology Inc
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    92
    Cell Signaling Technology Inc phospho specific stress activated protein kinases
    Effects of IGF-I on the phosphorylation of p44/p42 MAP kinase, p38 MAP kinase, SAPK/JNK, Akt and p70 S6 kinase in MC3T3-E1 cells. The cells were stimulated by 10 nM IGF-I for the indicated periods. Western blot analysis was performed using antibodies against <t>phospho-p44/p42</t> MAP kinase, phospho-p38 MAP kinase, phospho-SAPK/JNK, phospho-p70 S6 kinase, phospho-Akt and GAPDH. IGF-I, insulin-like growth factor-I; MAP, <t>mitogen-activated</t> <t>protein;</t> SAPK/JNK, <t>stress-activated</t> protein kinase/c-Jun N-terminal kinase.
    Phospho Specific Stress Activated Protein Kinases, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Ischemia/reperfusion-mediated activation of MAPKs in the lung. PKC β +/+ and PKC β –/– mice underwent the indicated period of left-lung I/R. Animals were sacrificed and protein extracts from the I/R and uninstrumented lung were prepared and subjected to SDS-PAGE (12%, 50 ∝g of protein/lane). Immunoblotting with phospho-p44/42 MAPK antibody or total p44/42 MAPK antibody ( B ), phospho-p38 MAPK antibody, or total p38 MAPK antibody ( I ), and phospho-SAPK/JNK antibody or total SAPK/JNK antibody ( J ) was performed. Data are shown as mean ± SEM of five experiments. Immunohistochemical analysis of phospho-p44/42 expression in murine lung from uninstrumented ( B ) or I/R ( C ) PKC β +/+ mice, and from uninstrumented ( D ) or I/R ( E ) PKC β –/– mice was performed. Scale bar: 50 ∝m. I/R lungs from PKC β +/+ mice were subjected to immunofluorescence microscopy and double stained with an anti–phospho-p44/42 antibody (red) ( F ) and an anti-macrophage antibody (F4/80, green) ( G ). The merging of F (pERK1/2) and G (F4/80) is shown in H . Arrows in F and arrowheads in G indicate dually stained cells Original magnification in F – H , ∞1,000. * P

    Journal: Journal of Clinical Investigation

    Article Title: PKC? regulates ischemia/reperfusion injury in the lung

    doi: 10.1172/JCI200419225

    Figure Lengend Snippet: Ischemia/reperfusion-mediated activation of MAPKs in the lung. PKC β +/+ and PKC β –/– mice underwent the indicated period of left-lung I/R. Animals were sacrificed and protein extracts from the I/R and uninstrumented lung were prepared and subjected to SDS-PAGE (12%, 50 ∝g of protein/lane). Immunoblotting with phospho-p44/42 MAPK antibody or total p44/42 MAPK antibody ( B ), phospho-p38 MAPK antibody, or total p38 MAPK antibody ( I ), and phospho-SAPK/JNK antibody or total SAPK/JNK antibody ( J ) was performed. Data are shown as mean ± SEM of five experiments. Immunohistochemical analysis of phospho-p44/42 expression in murine lung from uninstrumented ( B ) or I/R ( C ) PKC β +/+ mice, and from uninstrumented ( D ) or I/R ( E ) PKC β –/– mice was performed. Scale bar: 50 ∝m. I/R lungs from PKC β +/+ mice were subjected to immunofluorescence microscopy and double stained with an anti–phospho-p44/42 antibody (red) ( F ) and an anti-macrophage antibody (F4/80, green) ( G ). The merging of F (pERK1/2) and G (F4/80) is shown in H . Arrows in F and arrowheads in G indicate dually stained cells Original magnification in F – H , ∞1,000. * P

    Article Snippet: To detect MAPKs, each blot was incubated with one of the following antibodies as the first antibody for the reaction: anti–phospho-p44/42 IgG, anti–total-p44/42 IgG, anti–phospho-SAPK/JNK IgG, anti–total-SAPK/JNK IgG, anti-phospho-p38 IgG, or anti–total-p38 IgG (Cell Signaling Technology Inc.).

    Techniques: Activation Assay, Mouse Assay, SDS Page, Immunohistochemistry, Expressing, Immunofluorescence, Microscopy, Staining

    CFM-4.16 stimulates apoptosis in parental and TKI-resistant NSCLC cells in part by upregulating pro-apoptotic CARP-1 and activating SAPKs ( A – C ) Indicated parental and TKI-resistant NSCLC cells were either untreated (Control), treated with TKI, CFM-4, or CFM-4.16 for noted dose and time. Cell lysates were analyzed by Western blotting (WB) as in Methods for levels of CARP-1, cyclin B1, cleaved PARP and caspase-8, and activation (phosphorylation) of pro-apoptotic p38 and JNK1/2 SAPKs. The western blot membranes were subsequently probed with anti-actin or α-tubulin antibodies to assess equal loading. The presence of respective protein is indicated by an arrowhead on the left side of each blot. Approximate location of various molecular weight markers is indicated on the right side of each blot. kDa, kilodalton.

    Journal: Oncotarget

    Article Title: A CARP-1 functional mimetic compound is synergistic with BRAF-targeting in non-small cell lung cancers

    doi: 10.18632/oncotarget.25671

    Figure Lengend Snippet: CFM-4.16 stimulates apoptosis in parental and TKI-resistant NSCLC cells in part by upregulating pro-apoptotic CARP-1 and activating SAPKs ( A – C ) Indicated parental and TKI-resistant NSCLC cells were either untreated (Control), treated with TKI, CFM-4, or CFM-4.16 for noted dose and time. Cell lysates were analyzed by Western blotting (WB) as in Methods for levels of CARP-1, cyclin B1, cleaved PARP and caspase-8, and activation (phosphorylation) of pro-apoptotic p38 and JNK1/2 SAPKs. The western blot membranes were subsequently probed with anti-actin or α-tubulin antibodies to assess equal loading. The presence of respective protein is indicated by an arrowhead on the left side of each blot. Approximate location of various molecular weight markers is indicated on the right side of each blot. kDa, kilodalton.

    Article Snippet: We purchased antibodies for α-tubulin (rabbit polyclonal), Cyclin B1 (V152, mouse monoclonal), Cleaved Caspase-8 (IC12, mouse monoclonal), cleaved PARP (Asp214; mouse monoclonal), RIPK1 (Cat#3493; rabbit monoclonal), phospho (T180/Y182) and total p38α/β, phospho (T183/Y185) and total JNK1/2 SAPKs, total and phospho-STAT3 (Y705), total and phospho-MET (Y1234/1235), total and phospho (Y416)-Src, total and phospho-AKT (S473) and T(308), total and phospho (T389)-p70S6K, total and phospho (S2448)-mToR1, total and phospho-(Y1068) EGFR, total and phospho-(S536) p65/RelA NF-κB subunit, total and phospho (S445) B-Raf from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Western Blot, Activation Assay, Molecular Weight

    Analysis of signaling pathways involved in rLsa21-mediated cytokine production in mouse macrophages. ( A ) RAW264.7 cells were pretreated for 30 min at 37 °C with anti-TLR2 (30 μg/ml), anti-TLR4 (30 μg/ml), or an isotype control Ab (30 μg/ml) before stimulation with rLsa21 (5 μg/ml) for 30 min. Levels of phosphorylated p38, JNK, and ERK1/2 induced by rLsa21 and degraded IκBα were analyzed by western blot using anti–phospho-p38 (p-p38), or anti–phospho-ERK1/2 (p-ERK1/2), anti–phospho-JNK (p-JNK) and anti-mouse IκBα as well as a specific control Ab for each of the unphosphorylated kinases. Data are representative of those obtained in three independent experiments. ( B ) TLR2 −/− , TLR4 −/− or TLR2 −/− /TLR4 −/− DKO macrophages cell lines were stimulated with rLsa21 (5 μg/ml) for 30 min. Levels of phosphorylated p38, JNK, and ERK1/2 induced by rLsa21 were analyzed by western blot as described in materials and methods. ( C ) RAW 264.7 cells were pretreated for 30 min with NF-kB inhibitor (SN50; 20 μM), JNK inhibitor (SP600125; 40 μM) or p38MAPK inhibitor (SB203580; 30 μM) and then stimulated with rLsa21 (2 μg/ml). After incubation, supernatants were harvested and levels of IL-6 and TNF-α were measured by ELISA. Data are representative of three different experiments. (*Indicates P

    Journal: Scientific Reports

    Article Title: Leptospira surface adhesin (Lsa21) induces Toll like receptor 2 and 4 mediated inflammatory responses in macrophages

    doi: 10.1038/srep39530

    Figure Lengend Snippet: Analysis of signaling pathways involved in rLsa21-mediated cytokine production in mouse macrophages. ( A ) RAW264.7 cells were pretreated for 30 min at 37 °C with anti-TLR2 (30 μg/ml), anti-TLR4 (30 μg/ml), or an isotype control Ab (30 μg/ml) before stimulation with rLsa21 (5 μg/ml) for 30 min. Levels of phosphorylated p38, JNK, and ERK1/2 induced by rLsa21 and degraded IκBα were analyzed by western blot using anti–phospho-p38 (p-p38), or anti–phospho-ERK1/2 (p-ERK1/2), anti–phospho-JNK (p-JNK) and anti-mouse IκBα as well as a specific control Ab for each of the unphosphorylated kinases. Data are representative of those obtained in three independent experiments. ( B ) TLR2 −/− , TLR4 −/− or TLR2 −/− /TLR4 −/− DKO macrophages cell lines were stimulated with rLsa21 (5 μg/ml) for 30 min. Levels of phosphorylated p38, JNK, and ERK1/2 induced by rLsa21 were analyzed by western blot as described in materials and methods. ( C ) RAW 264.7 cells were pretreated for 30 min with NF-kB inhibitor (SN50; 20 μM), JNK inhibitor (SP600125; 40 μM) or p38MAPK inhibitor (SB203580; 30 μM) and then stimulated with rLsa21 (2 μg/ml). After incubation, supernatants were harvested and levels of IL-6 and TNF-α were measured by ELISA. Data are representative of three different experiments. (*Indicates P

    Article Snippet: After blocking with 5% nonfat milk in PBS (pH 7.4, 0.05% (v/v) Tween-20), the membranes were incubated overnight at 4 °C with primary Abs, including rabbit anti-mouse-p38, anti–mouse phospho-p38, anti-mouse-ERK2, anti–mouse phospho-ERK1/2, anti–mouse stress-activated protein kinase/JNK, anti–mouse phospho–stress-activated protein kinase/JNK, anti-mouse IκBα (Cell Signaling Technology, Beverly, MA), according to the manufacturer’s instructions.

    Techniques: Western Blot, Incubation, Enzyme-linked Immunosorbent Assay

    Effects of IGF-I on the phosphorylation of p44/p42 MAP kinase, p38 MAP kinase, SAPK/JNK, Akt and p70 S6 kinase in MC3T3-E1 cells. The cells were stimulated by 10 nM IGF-I for the indicated periods. Western blot analysis was performed using antibodies against phospho-p44/p42 MAP kinase, phospho-p38 MAP kinase, phospho-SAPK/JNK, phospho-p70 S6 kinase, phospho-Akt and GAPDH. IGF-I, insulin-like growth factor-I; MAP, mitogen-activated protein; SAPK/JNK, stress-activated protein kinase/c-Jun N-terminal kinase.

    Journal: Biomedical Reports

    Article Title: Repression of IGF-I-induced osteoblast migration by (−)-epigallocatechin gallate through p44/p42 MAP kinase signaling

    doi: 10.3892/br.2018.1140

    Figure Lengend Snippet: Effects of IGF-I on the phosphorylation of p44/p42 MAP kinase, p38 MAP kinase, SAPK/JNK, Akt and p70 S6 kinase in MC3T3-E1 cells. The cells were stimulated by 10 nM IGF-I for the indicated periods. Western blot analysis was performed using antibodies against phospho-p44/p42 MAP kinase, phospho-p38 MAP kinase, phospho-SAPK/JNK, phospho-p70 S6 kinase, phospho-Akt and GAPDH. IGF-I, insulin-like growth factor-I; MAP, mitogen-activated protein; SAPK/JNK, stress-activated protein kinase/c-Jun N-terminal kinase.

    Article Snippet: Phospho-specific p44/p42 MAP kinase antibody (cat. no. 9101), p44/p42 MAP kinase antibody (cat. no. 9102), phospho-specific p38 MAP kinase antibody (cat. no. 4511), phospho-specific stress-activated protein kinases (SAPK)/c-Jun N-terminal kinase (JNK) antibody (cat. no. 9251), phospho-specific p70 S6 kinase antibody (cat. no. 9205), phospho-specific Akt antibody (cat. no. 9275) and Akt antibody (cat. no. 9272) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Western Blot