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  • 99
    Name:
    p44 42 MAPK Erk1 2 Antibody
    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:
    9102
    Price:
    None
    Category:
    Primary Antibodies
    Source:
    Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to a sequence in the C-terminus of rat p44 MAP Kinase. Antibodies are purified by protein A and peptide affinity chromatography.
    Reactivity:
    Human Mouse Rat Hamster Monkey Mink Zebrafish Bovine Pig S cerevisiae
    Applications:
    Western Blot, Immunoprecipitation, Immunohistochemistry
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc anti erk
    Pazopanib induces autophagic flux and ERK1/2 phosphorylation in BC cells. ( A ) Lysates from 5637 and J82 cells treated with 20 μ M pazopanib and 50 n M bafilomycin A1, alone or in combination, for 48 h were separated on 14% or 10% SDS–PAGE and probed with <t>anti-LC3</t> or anti-p62 and anti-GAPDH Abs, respectively. ( B ) Lysates from 5637 and J82 cells untreated or treated with 20 μ M pazopanib at different times were separated on 12% SDS–PAGE and probed with anti-pERK or <t>anti-ERK</t> Abs. Densitometric analysis, expressed as the mean±s.d., was performed by comparing treated with untreated cells, * 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/anti erk/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti erk - by Bioz Stars, 2021-06
    99/100 stars

    Images

    1) Product Images from "Pazopanib and sunitinib trigger autophagic and non-autophagic death of bladder tumour cells"

    Article Title: Pazopanib and sunitinib trigger autophagic and non-autophagic death of bladder tumour cells

    Journal: British Journal of Cancer

    doi: 10.1038/bjc.2013.420

    Pazopanib induces autophagic flux and ERK1/2 phosphorylation in BC cells. ( A ) Lysates from 5637 and J82 cells treated with 20 μ M pazopanib and 50 n M bafilomycin A1, alone or in combination, for 48 h were separated on 14% or 10% SDS–PAGE and probed with anti-LC3 or anti-p62 and anti-GAPDH Abs, respectively. ( B ) Lysates from 5637 and J82 cells untreated or treated with 20 μ M pazopanib at different times were separated on 12% SDS–PAGE and probed with anti-pERK or anti-ERK Abs. Densitometric analysis, expressed as the mean±s.d., was performed by comparing treated with untreated cells, * P
    Figure Legend Snippet: Pazopanib induces autophagic flux and ERK1/2 phosphorylation in BC cells. ( A ) Lysates from 5637 and J82 cells treated with 20 μ M pazopanib and 50 n M bafilomycin A1, alone or in combination, for 48 h were separated on 14% or 10% SDS–PAGE and probed with anti-LC3 or anti-p62 and anti-GAPDH Abs, respectively. ( B ) Lysates from 5637 and J82 cells untreated or treated with 20 μ M pazopanib at different times were separated on 12% SDS–PAGE and probed with anti-pERK or anti-ERK Abs. Densitometric analysis, expressed as the mean±s.d., was performed by comparing treated with untreated cells, * P

    Techniques Used: SDS Page

    2) Product Images from "SV40 Polyomavirus Activates the Ras-MAPK Signaling Pathway for Vacuolization, Cell Death, and Virus Release"

    Article Title: SV40 Polyomavirus Activates the Ras-MAPK Signaling Pathway for Vacuolization, Cell Death, and Virus Release

    Journal: Viruses

    doi: 10.3390/v12101128

    MAP kinase components are required for efficient vacuolization, cell lysis, and virus release. ( a ) Inhibitors SP600125, Selumetinib, and SB203580 inhibit SV40-induced phosphorylation of JNK, ERK, and p38, respectively. CV-1 cells were infected at an MOI of 10. Inhibitor treatment was started at 12 h.p.i., and immunoblotting was performed on extracts prepared 48 h.p.i; ( b ) Bright-field images of CV-1 cells 48 h.p.i. after infection with SV40: CV-1 cells were infected at an MOI of 10 and treated with inhibitors against JNK, ERK, and p38 or DMSO vehicle at 12 h.p.i.. Scale bar, 20 µm; ( c ) The number of vacuolated cells two days after infection was quantified and normalized to DMSO-treated control cells. *** p
    Figure Legend Snippet: MAP kinase components are required for efficient vacuolization, cell lysis, and virus release. ( a ) Inhibitors SP600125, Selumetinib, and SB203580 inhibit SV40-induced phosphorylation of JNK, ERK, and p38, respectively. CV-1 cells were infected at an MOI of 10. Inhibitor treatment was started at 12 h.p.i., and immunoblotting was performed on extracts prepared 48 h.p.i; ( b ) Bright-field images of CV-1 cells 48 h.p.i. after infection with SV40: CV-1 cells were infected at an MOI of 10 and treated with inhibitors against JNK, ERK, and p38 or DMSO vehicle at 12 h.p.i.. Scale bar, 20 µm; ( c ) The number of vacuolated cells two days after infection was quantified and normalized to DMSO-treated control cells. *** p

    Techniques Used: Lysis, Infection

    SV40 infection activates intracellular signaling pathways at late times after infection. ( a ) Western blot analysis of phosphorylated and total p38, JNK, and ERK in mock-infected or SV40-infected CV-1 cells 48 h.p.i.: VP1 and beta-actin expression are shown as controls; ( b ) Western blot analysis of CV-1 cells over the time course of SV40 infection: Samples harvested at the indicated h.p.i. were analyzed for phosphorylated p38, JNK, ERK, MKK4, ATF2, and c-Jun as well as for beta-actin and VP1 expression.
    Figure Legend Snippet: SV40 infection activates intracellular signaling pathways at late times after infection. ( a ) Western blot analysis of phosphorylated and total p38, JNK, and ERK in mock-infected or SV40-infected CV-1 cells 48 h.p.i.: VP1 and beta-actin expression are shown as controls; ( b ) Western blot analysis of CV-1 cells over the time course of SV40 infection: Samples harvested at the indicated h.p.i. were analyzed for phosphorylated p38, JNK, ERK, MKK4, ATF2, and c-Jun as well as for beta-actin and VP1 expression.

    Techniques Used: Infection, Western Blot, Expressing

    3) Product Images from "ABCA7 Deficiency Accelerates Amyloid-β Generation and Alzheimer's Neuronal Pathology"

    Article Title: ABCA7 Deficiency Accelerates Amyloid-β Generation and Alzheimer's Neuronal Pathology

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3757-15.2016

    Possible pathways for ABCA7-related LOAD pathogenesis. ABCA7 dysfunctions likely facilitate Aβ production in neurons by activating the SREBP2-BACE1 pathway directly and/or by modifying the brain lipid profile indirectly. Furthermore, ER stress is also induced by ABCA7 loss of function in neurons, which activates the PERK-eIF2α pathway, causing cognitive declines and further activation of the SREBP2-BACE1 pathway. Excessive activation of the ERK signaling pathway caused by ABCA7 dysfunctions may also contribute directly to AD pathogenesis.
    Figure Legend Snippet: Possible pathways for ABCA7-related LOAD pathogenesis. ABCA7 dysfunctions likely facilitate Aβ production in neurons by activating the SREBP2-BACE1 pathway directly and/or by modifying the brain lipid profile indirectly. Furthermore, ER stress is also induced by ABCA7 loss of function in neurons, which activates the PERK-eIF2α pathway, causing cognitive declines and further activation of the SREBP2-BACE1 pathway. Excessive activation of the ERK signaling pathway caused by ABCA7 dysfunctions may also contribute directly to AD pathogenesis.

    Techniques Used: Activation Assay

    4) Product Images from "Cyp1b1 expression impacts the angiogenic and inflammatory properties of liver sinusoidal endothelial cells"

    Article Title: Cyp1b1 expression impacts the angiogenic and inflammatory properties of liver sinusoidal endothelial cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0206756

    Sustained activation of AKT and ERKs signaling pathways in Cyp1b1-/- LSEC. The activation of various signaling pathways were examined by Western blot analysis of cell lysates prepared from Cyp1b1 +/+ and Cyp1b1-/- LSEC as detailed in Methods. Please note an increase in the levels of phosphorylated AKT (p-AKT) and ERK (p-ERK), and a decrease in p-JNK in Cyp1b1-/- cells. The levels of phosphorylated Src (p-Src416 and p-Src527) and MAPK p38 (p-p38) were not affected. Similar levels of total proteins were observed in both cell types. These experiments were repeated with two isolations of LSEC with similar results.
    Figure Legend Snippet: Sustained activation of AKT and ERKs signaling pathways in Cyp1b1-/- LSEC. The activation of various signaling pathways were examined by Western blot analysis of cell lysates prepared from Cyp1b1 +/+ and Cyp1b1-/- LSEC as detailed in Methods. Please note an increase in the levels of phosphorylated AKT (p-AKT) and ERK (p-ERK), and a decrease in p-JNK in Cyp1b1-/- cells. The levels of phosphorylated Src (p-Src416 and p-Src527) and MAPK p38 (p-p38) were not affected. Similar levels of total proteins were observed in both cell types. These experiments were repeated with two isolations of LSEC with similar results.

    Techniques Used: Activation Assay, Western Blot

    5) Product Images from "Autotaxin-LPA signaling contributes to obesity-induced insulin resistance in muscle and impairs mitochondrial metabolism"

    Article Title: Autotaxin-LPA signaling contributes to obesity-induced insulin resistance in muscle and impairs mitochondrial metabolism

    Journal: Journal of Lipid Research

    doi: 10.1194/jlr.M082008

    LPA impairs insulin signaling and exacerbates palmitate-induced insulin resistance in C2C12 myotubes. Immunoblot and densitometric analysis of AKT phosphorylation at S 473 (A, B), ERK phosphorylation at T 202/ Y 204 (A, C), and JNK phosphorylation at T T183/ Y 185 (A, D) in C2C12 myotubes incubated in the absence or presence of 0.8 mM palmitate and 0, 1, or 10 μM LPA for 18 h, followed by stimulation with 20 nM insulin for 15 min (n = 6). B–D: Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test. * P
    Figure Legend Snippet: LPA impairs insulin signaling and exacerbates palmitate-induced insulin resistance in C2C12 myotubes. Immunoblot and densitometric analysis of AKT phosphorylation at S 473 (A, B), ERK phosphorylation at T 202/ Y 204 (A, C), and JNK phosphorylation at T T183/ Y 185 (A, D) in C2C12 myotubes incubated in the absence or presence of 0.8 mM palmitate and 0, 1, or 10 μM LPA for 18 h, followed by stimulation with 20 nM insulin for 15 min (n = 6). B–D: Statistical analysis was performed using a two-way ANOVA followed by a Tukey’s multiple comparison test. * P

    Techniques Used: Incubation

    6) Product Images from "Targeting hyperactivation of the AKT survival pathway to overcome therapy resistance of melanoma brain metastases"

    Article Title: Targeting hyperactivation of the AKT survival pathway to overcome therapy resistance of melanoma brain metastases

    Journal: Cancer Medicine

    doi: 10.1002/cam4.50

    (A–E) Brain and lung metastases from a melanoma patient were stained to detect the melanocytic marker Melan A, total and activated ERK (p-ERK), total and activated AKT (p-AKT), and PTEN; 40×: 40-fold magnification, 200×: 200-fold magnification. (F) Brain metastasis from a melanoma patient was stained for p-AKT (red) and HMB-45 (brown) to detect melanoma cells, 200×: 200-fold magnification. The insert shows colocalization of p-AKT (red) with HMB-45 (brown), 630-fold magnification. (G) Brain metastasis from a melanoma patient was stained for p-AKT (red) and glial fibrillary acidic protein (GFAP) (brown) to detect glial cells, 200×: 200-fold magnification. GFAP does not colocalize with p-AKT. (H) Brain metastasis from a melanoma patient was stained for p-AKT (red) and CD14 (brown) to detect monocytes/macrophages, 200×: 200-fold magnification. CD14 does not colocalize with p-AKT.
    Figure Legend Snippet: (A–E) Brain and lung metastases from a melanoma patient were stained to detect the melanocytic marker Melan A, total and activated ERK (p-ERK), total and activated AKT (p-AKT), and PTEN; 40×: 40-fold magnification, 200×: 200-fold magnification. (F) Brain metastasis from a melanoma patient was stained for p-AKT (red) and HMB-45 (brown) to detect melanoma cells, 200×: 200-fold magnification. The insert shows colocalization of p-AKT (red) with HMB-45 (brown), 630-fold magnification. (G) Brain metastasis from a melanoma patient was stained for p-AKT (red) and glial fibrillary acidic protein (GFAP) (brown) to detect glial cells, 200×: 200-fold magnification. GFAP does not colocalize with p-AKT. (H) Brain metastasis from a melanoma patient was stained for p-AKT (red) and CD14 (brown) to detect monocytes/macrophages, 200×: 200-fold magnification. CD14 does not colocalize with p-AKT.

    Techniques Used: Staining, Marker

    7) Product Images from "Anti-inflammatory effects of indirubin derivatives on influenza A virus-infected human pulmonary microvascular endothelial cells"

    Article Title: Anti-inflammatory effects of indirubin derivatives on influenza A virus-infected human pulmonary microvascular endothelial cells

    Journal: Scientific Reports

    doi: 10.1038/srep18941

    Infection of H9N2 on human pulmonary endothelial cells induces rapid phosphorylation of p38, JNK, and STAT3. HPMECs were infected with H9N2 at 2 MOI. Total cell lysates were harvested at the indicated time (0.25, 0.5 and 1 h after addition of virus, and 0.25, 0.5, 1, 6 and 24 h.p.i.). Phosphorylation and total p38, JNK, ERK and STAT3 were detected by Western blot analysis with specific antibodies. ( A ) Representative image from three independent experiments. ( B ) Quantitative analysis of Western blotting. Values are presented as fold-change of phosphorylated p38, JNK, ERK or STAT3 normalized to the corresponding total form and then compared with mock-infected cells. The values are presented as mean ± S.D. from three independent experiments. * p
    Figure Legend Snippet: Infection of H9N2 on human pulmonary endothelial cells induces rapid phosphorylation of p38, JNK, and STAT3. HPMECs were infected with H9N2 at 2 MOI. Total cell lysates were harvested at the indicated time (0.25, 0.5 and 1 h after addition of virus, and 0.25, 0.5, 1, 6 and 24 h.p.i.). Phosphorylation and total p38, JNK, ERK and STAT3 were detected by Western blot analysis with specific antibodies. ( A ) Representative image from three independent experiments. ( B ) Quantitative analysis of Western blotting. Values are presented as fold-change of phosphorylated p38, JNK, ERK or STAT3 normalized to the corresponding total form and then compared with mock-infected cells. The values are presented as mean ± S.D. from three independent experiments. * p

    Techniques Used: Infection, Western Blot

    8) Product Images from "Downregulation of KIAA1199 by miR‐486‐5p suppresses tumorigenesis in lung cancer, et al. Downregulation of KIAA1199 by miR‐486‐5p suppresses tumorigenesis in lung cancer"

    Article Title: Downregulation of KIAA1199 by miR‐486‐5p suppresses tumorigenesis in lung cancer, et al. Downregulation of KIAA1199 by miR‐486‐5p suppresses tumorigenesis in lung cancer

    Journal: Cancer Medicine

    doi: 10.1002/cam4.3210

    A, A549 and SPC‐A1 cells were transfected with miR‐486‐5p mimics or inhibitor for 72 hours. The levels of pEGFR, EGFR, pErk, Erk, pAKT, AKT, pSmad3, Smad3, and other EMT markers were analyzed by western blotting. B, A diagram shows the mechanism of which miR‐485‐5p interacts with KIAA1199 and their control of EGFR‐related signaling pathways
    Figure Legend Snippet: A, A549 and SPC‐A1 cells were transfected with miR‐486‐5p mimics or inhibitor for 72 hours. The levels of pEGFR, EGFR, pErk, Erk, pAKT, AKT, pSmad3, Smad3, and other EMT markers were analyzed by western blotting. B, A diagram shows the mechanism of which miR‐485‐5p interacts with KIAA1199 and their control of EGFR‐related signaling pathways

    Techniques Used: Transfection, Western Blot

    9) Product Images from "GSTM3 and GSTP1: novel players driving tumor progression in cervical cancer"

    Article Title: GSTM3 and GSTP1: novel players driving tumor progression in cervical cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.24796

    ( A ) Cytoscape interaction network representing GSTM3–prey interactions. GSTM3–prey interactions were visualized by network edges. This analysis was performed to obtain the protein-protein interactions reported by the SysBiomics databases, in which we observed that TRAF6 interact with GSTM3. ( B ) Co-immunoprecipitation of GSTM3 and TRAF6. ( C ) Western blotting was performed for TRAF6, ERK, p-ERK, pERK, NF-κB, pNF-κB, IKBα, pIKBα, p38, pp38, JNK, pJNK, and TLR4 in the protein extracts from tumors xenotransplants in female mice. ( D ) Proportional Venn diagram of secreted proteins of CC cell lines with 264 common proteins. ( E ) Two proteins were identified in secreted proteins that can activate the TLR4 signal-pathway expressed in vitro in HSP60 and HPS70 ( F ) Western blot of TRL4 activators HSP70 and HSP60 in the proteins secreted by CC tumors, HSP60 was expressed in SiHa and HeLa tumors at day 50, and HSP70 protein expressed in SiHa tumors at 43 days and Hela tumors at 30 and 50 days.
    Figure Legend Snippet: ( A ) Cytoscape interaction network representing GSTM3–prey interactions. GSTM3–prey interactions were visualized by network edges. This analysis was performed to obtain the protein-protein interactions reported by the SysBiomics databases, in which we observed that TRAF6 interact with GSTM3. ( B ) Co-immunoprecipitation of GSTM3 and TRAF6. ( C ) Western blotting was performed for TRAF6, ERK, p-ERK, pERK, NF-κB, pNF-κB, IKBα, pIKBα, p38, pp38, JNK, pJNK, and TLR4 in the protein extracts from tumors xenotransplants in female mice. ( D ) Proportional Venn diagram of secreted proteins of CC cell lines with 264 common proteins. ( E ) Two proteins were identified in secreted proteins that can activate the TLR4 signal-pathway expressed in vitro in HSP60 and HPS70 ( F ) Western blot of TRL4 activators HSP70 and HSP60 in the proteins secreted by CC tumors, HSP60 was expressed in SiHa and HeLa tumors at day 50, and HSP70 protein expressed in SiHa tumors at 43 days and Hela tumors at 30 and 50 days.

    Techniques Used: Immunoprecipitation, Western Blot, Mouse Assay, In Vitro

    10) Product Images from "Deactivation of STAT6 through Serine 707 Phosphorylation by JNK *"

    Article Title: Deactivation of STAT6 through Serine 707 Phosphorylation by JNK *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.168435

    Specificity of kinases for Ser-707 phosphorylation of STAT6. A , purified GST-STAT6WT was incubated in the presence or absence of purified active JNK ( lanes 1 and 2 ), p38 ( lanes 3 and 4 ), ERK ( lanes 5 and 6 ), or AKT ( lanes 7 and 8 ). Products were separated on a 10% SDS-polyacrylamide gel, and analyzed by Coomassie staining ( upper panel ) or autoradiogram ( lower panel ). B , effects of JNK, p38, ERK, or AKT knockdown on Ser-707 phosphorylation of STAT6. siRNAs were transfected into HeLa cells, which were incubated with DMSO (control) or anisomycin (500 ng/ml) for 45 min. The levels of Ser-707 phosphorylation of STAT6 were observed as a mobility shift in Western blots. The arrow indicates a shifted band.
    Figure Legend Snippet: Specificity of kinases for Ser-707 phosphorylation of STAT6. A , purified GST-STAT6WT was incubated in the presence or absence of purified active JNK ( lanes 1 and 2 ), p38 ( lanes 3 and 4 ), ERK ( lanes 5 and 6 ), or AKT ( lanes 7 and 8 ). Products were separated on a 10% SDS-polyacrylamide gel, and analyzed by Coomassie staining ( upper panel ) or autoradiogram ( lower panel ). B , effects of JNK, p38, ERK, or AKT knockdown on Ser-707 phosphorylation of STAT6. siRNAs were transfected into HeLa cells, which were incubated with DMSO (control) or anisomycin (500 ng/ml) for 45 min. The levels of Ser-707 phosphorylation of STAT6 were observed as a mobility shift in Western blots. The arrow indicates a shifted band.

    Techniques Used: Purification, Incubation, Staining, Transfection, Mobility Shift, Western Blot

    11) Product Images from "MiR-103 alleviates autophagy and apoptosis by regulating SOX2 in LPS-injured PC12 cells and SCI rats"

    Article Title: MiR-103 alleviates autophagy and apoptosis by regulating SOX2 in LPS-injured PC12 cells and SCI rats

    Journal: Iranian Journal of Basic Medical Sciences

    doi: 10.22038/ijbms.2018.25980.6392

    Proposed mechanisms of miR-103 effect on apoptosis and autophagy in LPS-injured PC12 cells and SCI rats. In pc12 cells, miR-103 enhances the expression of SOX2, and SOX2 further reduced LPS-induced cell apoptosis and autophagy. In addition, SOX2 inhibits the activation of ERK/MAPK and JAK/STAT3 pathway. In SCI rats, SCI induces the down-regulation of miR-103 and SOX2. miR-103 agomir promotes the expression of SOX2, and suppresses cell apoptosis and autophagy in SCI rats. * P
    Figure Legend Snippet: Proposed mechanisms of miR-103 effect on apoptosis and autophagy in LPS-injured PC12 cells and SCI rats. In pc12 cells, miR-103 enhances the expression of SOX2, and SOX2 further reduced LPS-induced cell apoptosis and autophagy. In addition, SOX2 inhibits the activation of ERK/MAPK and JAK/STAT3 pathway. In SCI rats, SCI induces the down-regulation of miR-103 and SOX2. miR-103 agomir promotes the expression of SOX2, and suppresses cell apoptosis and autophagy in SCI rats. * P

    Techniques Used: Expressing, Activation Assay

    MiR-103 inhibited MAPK/ERK pathway and JAK/STAT pathway through upregulation of SOX2. PC12 cells were transfected with miR-103 mimic and mimic control, or co-transfected with sh-SOX2. And cells were incubated with 5 μg/ml LPS for 12 hr. The protein expressions of core factors related with (A) MAPK/ERK signaling pathway and (B) JAK/STAT signaling pathways were measured by Western blot
    Figure Legend Snippet: MiR-103 inhibited MAPK/ERK pathway and JAK/STAT pathway through upregulation of SOX2. PC12 cells were transfected with miR-103 mimic and mimic control, or co-transfected with sh-SOX2. And cells were incubated with 5 μg/ml LPS for 12 hr. The protein expressions of core factors related with (A) MAPK/ERK signaling pathway and (B) JAK/STAT signaling pathways were measured by Western blot

    Techniques Used: Transfection, Incubation, Western Blot

    12) Product Images from "CTLA-4 positive breast cancer cells suppress dendritic cells maturation and function"

    Article Title: CTLA-4 positive breast cancer cells suppress dendritic cells maturation and function

    Journal: Oncotarget

    doi: 10.18632/oncotarget.14626

    The relevance of ERK and STAT3 activation by CTLA-4 in CTLA-4 + BCCs-treated DC and the direct inhibitory effects of CTLA-4 mAb on CTLA-4 + BCCs ( A ) ImDCs were cultured in medium (control), or with soluble CTLA-4 (CTLA-4), or coculture with CTLA-4 + BCCs alone (231) or in the presence of CTLA-4 mAb (Ab) for 1 hour. Then, treated cells were collected. Whole cell extracts were prepared and the protein levels of phosphorylated (p) and total ERK1/2 and STAT3 were determined by western blot. Right insets: the quantitative analysis of p-ERK/ERK, p-STAT3/STAT3 protein ratio, as measured by ImageJ analysis of band intensity. Values are expressed in arbitrary units. ( B ) CCK-8 assays showed the effects of CTLA-4 mAb on cell viability. ( C ) Flow cytometry assays showed the effects of CTLA-4 mAb on cell cycle progression. ( D )Annexin staining assays showed the effects of CTLA-4 mAb on cell apoptosis. Representative result from three independent experiments was shown. * P
    Figure Legend Snippet: The relevance of ERK and STAT3 activation by CTLA-4 in CTLA-4 + BCCs-treated DC and the direct inhibitory effects of CTLA-4 mAb on CTLA-4 + BCCs ( A ) ImDCs were cultured in medium (control), or with soluble CTLA-4 (CTLA-4), or coculture with CTLA-4 + BCCs alone (231) or in the presence of CTLA-4 mAb (Ab) for 1 hour. Then, treated cells were collected. Whole cell extracts were prepared and the protein levels of phosphorylated (p) and total ERK1/2 and STAT3 were determined by western blot. Right insets: the quantitative analysis of p-ERK/ERK, p-STAT3/STAT3 protein ratio, as measured by ImageJ analysis of band intensity. Values are expressed in arbitrary units. ( B ) CCK-8 assays showed the effects of CTLA-4 mAb on cell viability. ( C ) Flow cytometry assays showed the effects of CTLA-4 mAb on cell cycle progression. ( D )Annexin staining assays showed the effects of CTLA-4 mAb on cell apoptosis. Representative result from three independent experiments was shown. * P

    Techniques Used: Activation Assay, Cell Culture, Western Blot, CCK-8 Assay, Flow Cytometry, Cytometry, Staining

    13) Product Images from "The decay of Redox-stress Response Capacity is a substantive characteristic of aging: Revising the redox theory of aging"

    Article Title: The decay of Redox-stress Response Capacity is a substantive characteristic of aging: Revising the redox theory of aging

    Journal: Redox Biology

    doi: 10.1016/j.redox.2016.12.026

    Moderate ROS generation in young cells activates signaling pathways. (A) p-AKT, AKT, p-ERK and ERK protein levels in p34 and p42 cells in response to PQ treatment were assessed by performing Western blotting at the indicated times. p-AKT and p-ERK levels were normalized to AKT and ERK levels, respectively. (B) p-AKT, AKT, p-ERK and t-ERK protein levels in p34 and p42 cells treated with PQ for 12 h in the presence or absence of the antioxidant NAC (Sigma) were determined by performing Western blotting. p-AKT and p-ERK levels were normalized to AKT and ERK levels, respectively. (C) p-AMPKα, AMPKα, p-AMPKβ and AMPKβ protein levels in p34 and p42 cells in response to PQ treatment were assessed by performing Western blotting at the indicated times. p-AMPKα and p-AMPKβ levels were normalized to AMPKα and p-AMPKβ levels, respectively. (D) HIF-1α levels were determined by performing qRT-PCR on p34 and p42 cells under normoxia (21% O 2 ) or hypoxia (3% O 2 ). (E) HIF-1α protein levels were determined by performing Western blotting under normoxia or hypoxia and normalized to Actin levels.
    Figure Legend Snippet: Moderate ROS generation in young cells activates signaling pathways. (A) p-AKT, AKT, p-ERK and ERK protein levels in p34 and p42 cells in response to PQ treatment were assessed by performing Western blotting at the indicated times. p-AKT and p-ERK levels were normalized to AKT and ERK levels, respectively. (B) p-AKT, AKT, p-ERK and t-ERK protein levels in p34 and p42 cells treated with PQ for 12 h in the presence or absence of the antioxidant NAC (Sigma) were determined by performing Western blotting. p-AKT and p-ERK levels were normalized to AKT and ERK levels, respectively. (C) p-AMPKα, AMPKα, p-AMPKβ and AMPKβ protein levels in p34 and p42 cells in response to PQ treatment were assessed by performing Western blotting at the indicated times. p-AMPKα and p-AMPKβ levels were normalized to AMPKα and p-AMPKβ levels, respectively. (D) HIF-1α levels were determined by performing qRT-PCR on p34 and p42 cells under normoxia (21% O 2 ) or hypoxia (3% O 2 ). (E) HIF-1α protein levels were determined by performing Western blotting under normoxia or hypoxia and normalized to Actin levels.

    Techniques Used: Western Blot, Quantitative RT-PCR

    14) Product Images from "A Novel Monoclonal Antibody Targets Mucin1 and Attenuates Growth in Pancreatic Cancer Model"

    Article Title: A Novel Monoclonal Antibody Targets Mucin1 and Attenuates Growth in Pancreatic Cancer Model

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19072004

    Inhibition of EGF-induced ERK phosphorylation and cyclin D1 expression by anti-hMUC1 monoclonal antibody. ( a ) Capan-2 and ( b ) PANC-1 cells were pretreated with 10 µg/mL anti-hMUC1 monoclonal antibody (anti-hMUC1 Ab) for 6 h followed by stimulation with 10 ng/mL EGF for 5 min. Cell lysates were separated by SDS-PAGE and western blot analysis was performed with anti-phospho-ERK, anti-ERK and anti-β-actin antibodies. ( c ) Capan-2 cells and ( d ) PANC-1 cells were treated with 10 µg/mL anti-hMUC1 monoclonal antibody for 6 h followed by treatment with 10 ng/mL EGF and incubated for 24 h. Cell lysates were separated by SDS-PAGE and western blot analysis was performed with anti-cyclin D1 and anti-β-actin antibodies. These results are representatives of three independent experiments.
    Figure Legend Snippet: Inhibition of EGF-induced ERK phosphorylation and cyclin D1 expression by anti-hMUC1 monoclonal antibody. ( a ) Capan-2 and ( b ) PANC-1 cells were pretreated with 10 µg/mL anti-hMUC1 monoclonal antibody (anti-hMUC1 Ab) for 6 h followed by stimulation with 10 ng/mL EGF for 5 min. Cell lysates were separated by SDS-PAGE and western blot analysis was performed with anti-phospho-ERK, anti-ERK and anti-β-actin antibodies. ( c ) Capan-2 cells and ( d ) PANC-1 cells were treated with 10 µg/mL anti-hMUC1 monoclonal antibody for 6 h followed by treatment with 10 ng/mL EGF and incubated for 24 h. Cell lysates were separated by SDS-PAGE and western blot analysis was performed with anti-cyclin D1 and anti-β-actin antibodies. These results are representatives of three independent experiments.

    Techniques Used: Inhibition, Expressing, SDS Page, Western Blot, Incubation

    15) Product Images from "Role of calpain-mediated p53 truncation in semaphorin 3A-induced axonal growth regulation"

    Article Title: Role of calpain-mediated p53 truncation in semaphorin 3A-induced axonal growth regulation

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1008652107

    ERK and p38 MAPK activation is involved in p-p53 truncation and calpain activation. ( A ) Immunoblotting analysis of various proteins in cultured cortical neurons. Cortical neurons were treated on DIV4 with DMSO (lane 1), semaphorin 3A (lane 2), semaphorin
    Figure Legend Snippet: ERK and p38 MAPK activation is involved in p-p53 truncation and calpain activation. ( A ) Immunoblotting analysis of various proteins in cultured cortical neurons. Cortical neurons were treated on DIV4 with DMSO (lane 1), semaphorin 3A (lane 2), semaphorin

    Techniques Used: Activation Assay, Cell Culture

    Semaphorin 3A–induced growth cone collapse and p53 degradation are blocked by ERK and p38 inhibition. Cultured hippocampal neurons were treated on DIV 4 with DMSO or semaphorin 3A (SM3A) in the presence or absence of U0126 or SB203580 pretreatment
    Figure Legend Snippet: Semaphorin 3A–induced growth cone collapse and p53 degradation are blocked by ERK and p38 inhibition. Cultured hippocampal neurons were treated on DIV 4 with DMSO or semaphorin 3A (SM3A) in the presence or absence of U0126 or SB203580 pretreatment

    Techniques Used: Inhibition, Cell Culture

    16) Product Images from "The expression and clinical significance of HDGF in osteosarcoma"

    Article Title: The expression and clinical significance of HDGF in osteosarcoma

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S91708

    HDGF could stimulate AKT and MAPK signaling pathway. Notes: ( A ) HDGF concentration in medium and cell lysate of U2-OS was detected with ELISA method 48 hours after transfection with scrambled RNA, HDGF siRNA, or Lipofectamine 2000. ( B ) Recombinant HDGF effect on phosphorylation of AKT, ERK, and p38 in U2-OS cells. U2-OS cells were stimulated with 10 µg/mL HDGF for 0 to 180 minutes, then lysed and detected by primary antibodies of AKT, p-AKT-Ser473, ERK, p-ERK-Thr202/Tyr204, p38, p-p38-Thr180/Tyr182, and β-actin. ( C ) AKT inhibitor wortmannin was able to decrease HDGF-induced phosphorylation of AKT/ERK/p38. U2-OS cells were pre-incubated in 10 µM wortmannin for 30 minutes and then stimulated in recombinant HDGF for 30 minutes. Immunoblotting was used to detect the phosphorylation of AKT/ERK/p38. Abbreviations: ELISA, enzyme-linked immunosorbent assay; IB, immunoblotting; HDGF, hepatoma-derived growth factor; OS, osteosarcoma.
    Figure Legend Snippet: HDGF could stimulate AKT and MAPK signaling pathway. Notes: ( A ) HDGF concentration in medium and cell lysate of U2-OS was detected with ELISA method 48 hours after transfection with scrambled RNA, HDGF siRNA, or Lipofectamine 2000. ( B ) Recombinant HDGF effect on phosphorylation of AKT, ERK, and p38 in U2-OS cells. U2-OS cells were stimulated with 10 µg/mL HDGF for 0 to 180 minutes, then lysed and detected by primary antibodies of AKT, p-AKT-Ser473, ERK, p-ERK-Thr202/Tyr204, p38, p-p38-Thr180/Tyr182, and β-actin. ( C ) AKT inhibitor wortmannin was able to decrease HDGF-induced phosphorylation of AKT/ERK/p38. U2-OS cells were pre-incubated in 10 µM wortmannin for 30 minutes and then stimulated in recombinant HDGF for 30 minutes. Immunoblotting was used to detect the phosphorylation of AKT/ERK/p38. Abbreviations: ELISA, enzyme-linked immunosorbent assay; IB, immunoblotting; HDGF, hepatoma-derived growth factor; OS, osteosarcoma.

    Techniques Used: Concentration Assay, Enzyme-linked Immunosorbent Assay, Transfection, Recombinant, Incubation, Derivative Assay

    17) Product Images from "Effects of the Oncoprotein PAX3-FOXO1 on Modulation of Exosomes Function and Protein Content: Implications on Oxidative Stress Protection and Enhanced Plasticity"

    Article Title: Effects of the Oncoprotein PAX3-FOXO1 on Modulation of Exosomes Function and Protein Content: Implications on Oxidative Stress Protection and Enhanced Plasticity

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2020.01784

    C2C12-P3F-derived exosomes promote angiogenesis and induce stemness of recipient cells. (A) Western blot for the indicated proteins in MEFs treated with Exo-free (EF) media, 10X Ctrl-C2C12-derived exosomes, and 10X P3F-C2C12-derived exosomes. GAPDH is used as a loading control. Histograms presenting the levels of phosphorylation of ERK and 4EBP1 compared to Exo-free condition, quantified by ImageJ software from n = 3 different western blots. (B) Zymography image of digested regions showing MMP2 activity in MEFs treated with either Ctrl-C2C12-derived exosomes or P3F-C2C12-derived exosomes for 72 h. Relative MMP-2 activity from 3 independent experiments was quantified by ImageJ software and presented by a histogram. (C) Representative phase contrast photomicrographs of endothelial tube formation of HUVECs cultured on Matrigel with either Exo-Free medium (EF), 1X, or 10X C2C12-MSCV-derived exosomes or C2C12-P3F-derived exosomes, as specified. Quantitation of the total number of nodes, total length and total mesh area of the different conditions are shown in the histogram, which represent the mean of 3 independent experiments, each performed in triplicate. (D,E) Representative phase contrast photomicrographs and histograms showing the ratio of the spheres formed by p53−/− MEFs (D) or C2C12 cells (E) treated with either Exo-Free medium (EF) or 10X C2C12-MSCV-derived exosomes or C2C12-P3F-derived exosomes, as specified. All data are reported as means of at least three independent experiments ± SD. Asterisks (*) denote a statistically significant difference ( p -value
    Figure Legend Snippet: C2C12-P3F-derived exosomes promote angiogenesis and induce stemness of recipient cells. (A) Western blot for the indicated proteins in MEFs treated with Exo-free (EF) media, 10X Ctrl-C2C12-derived exosomes, and 10X P3F-C2C12-derived exosomes. GAPDH is used as a loading control. Histograms presenting the levels of phosphorylation of ERK and 4EBP1 compared to Exo-free condition, quantified by ImageJ software from n = 3 different western blots. (B) Zymography image of digested regions showing MMP2 activity in MEFs treated with either Ctrl-C2C12-derived exosomes or P3F-C2C12-derived exosomes for 72 h. Relative MMP-2 activity from 3 independent experiments was quantified by ImageJ software and presented by a histogram. (C) Representative phase contrast photomicrographs of endothelial tube formation of HUVECs cultured on Matrigel with either Exo-Free medium (EF), 1X, or 10X C2C12-MSCV-derived exosomes or C2C12-P3F-derived exosomes, as specified. Quantitation of the total number of nodes, total length and total mesh area of the different conditions are shown in the histogram, which represent the mean of 3 independent experiments, each performed in triplicate. (D,E) Representative phase contrast photomicrographs and histograms showing the ratio of the spheres formed by p53−/− MEFs (D) or C2C12 cells (E) treated with either Exo-Free medium (EF) or 10X C2C12-MSCV-derived exosomes or C2C12-P3F-derived exosomes, as specified. All data are reported as means of at least three independent experiments ± SD. Asterisks (*) denote a statistically significant difference ( p -value

    Techniques Used: Derivative Assay, Western Blot, Software, Zymography, Activity Assay, Cell Culture, Quantitation Assay

    18) Product Images from "Pinin associates with prognosis of hepatocellular carcinoma through promoting cell proliferation and suppressing glucose deprivation-induced apoptosis"

    Article Title: Pinin associates with prognosis of hepatocellular carcinoma through promoting cell proliferation and suppressing glucose deprivation-induced apoptosis

    Journal: Oncotarget

    doi: 10.18632/oncotarget.9233

    Effects of ERK inactivation on Pinin modulated HCC apoptosis during glucose deprivation a. SNU449 and BEL7402 cells with or without knockdown of Pinin were cultured in present of glucose or not for the indicated periods. Cell lysates were analyzed with the indicated antibodies. GAPDH was detected as the loading control. b. Quantification of pERK levels relative to GAPDH is shown. c. Pinin was overexpressed in SNU449 and BEL7402 cells and then the cells were cultured in present of glucose or not for 24 h. Cell lysates were analyzed with the antibodies indicated. GAPDH was detected as the loading control. d. Pinin was overexpressed in SNU449 and BEL7402 cells and then the cells were cultured in present of glucose or not for 24h with or without U0126 treatment. Cell lysates were analyzed with the antibodies indicated. GAPDH was detected as the loading control. e-h. The percentage of cell apoptosis was analyzed by flow cytometry analysis in SNU449 and BEL7402 cells. (n=3, mean ± SD, t -test, ** P
    Figure Legend Snippet: Effects of ERK inactivation on Pinin modulated HCC apoptosis during glucose deprivation a. SNU449 and BEL7402 cells with or without knockdown of Pinin were cultured in present of glucose or not for the indicated periods. Cell lysates were analyzed with the indicated antibodies. GAPDH was detected as the loading control. b. Quantification of pERK levels relative to GAPDH is shown. c. Pinin was overexpressed in SNU449 and BEL7402 cells and then the cells were cultured in present of glucose or not for 24 h. Cell lysates were analyzed with the antibodies indicated. GAPDH was detected as the loading control. d. Pinin was overexpressed in SNU449 and BEL7402 cells and then the cells were cultured in present of glucose or not for 24h with or without U0126 treatment. Cell lysates were analyzed with the antibodies indicated. GAPDH was detected as the loading control. e-h. The percentage of cell apoptosis was analyzed by flow cytometry analysis in SNU449 and BEL7402 cells. (n=3, mean ± SD, t -test, ** P

    Techniques Used: Cell Culture, Flow Cytometry, Cytometry

    19) Product Images from "Anti-inflammatory effect of tribulusamide D isolated from Tribulus terrestris in lipopolysaccharide-stimulated RAW264.7 macrophages"

    Article Title: Anti-inflammatory effect of tribulusamide D isolated from Tribulus terrestris in lipopolysaccharide-stimulated RAW264.7 macrophages

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.7208

    Regulatory effect of tribulusamide D on NF-κB nuclear localization and phosphorylation of p38 mitogen-activated protein kinase in LPS-stimulated RAW 264.7 cells. Cells were treated with tribulusamide D (25–100 µM) for 1 h prior to LPS (0.5 µg/ml) stimulation for 30 min. (A) Cytosolic and nuclear extracts were analyzed via western blot analysis with anti-NF-κB p65, , anti-β-actin or anti-lamin B antibodies. (B) Cell lysates were subjected to western blot analysis with anti-p-ERK, anti-ERK, anti-p-JNK, anti-JNK, anti-p-p38 and anti-p38 antibodies. Results are representative of at least three independent replicates. NF-κB, nuclear factor-κB; LPS, lipopolysaccharide; p-, phospho-; JNK, c-Jun N-terminal kinase; ERK, extracellular signal-regulated kinase.
    Figure Legend Snippet: Regulatory effect of tribulusamide D on NF-κB nuclear localization and phosphorylation of p38 mitogen-activated protein kinase in LPS-stimulated RAW 264.7 cells. Cells were treated with tribulusamide D (25–100 µM) for 1 h prior to LPS (0.5 µg/ml) stimulation for 30 min. (A) Cytosolic and nuclear extracts were analyzed via western blot analysis with anti-NF-κB p65, , anti-β-actin or anti-lamin B antibodies. (B) Cell lysates were subjected to western blot analysis with anti-p-ERK, anti-ERK, anti-p-JNK, anti-JNK, anti-p-p38 and anti-p38 antibodies. Results are representative of at least three independent replicates. NF-κB, nuclear factor-κB; LPS, lipopolysaccharide; p-, phospho-; JNK, c-Jun N-terminal kinase; ERK, extracellular signal-regulated kinase.

    Techniques Used: Western Blot

    20) Product Images from "VPS35 regulates cell surface recycling and signaling of dopamine receptor D1"

    Article Title: VPS35 regulates cell surface recycling and signaling of dopamine receptor D1

    Journal: Neurobiology of aging

    doi: 10.1016/j.neurobiolaging.2016.05.016

    VPS35 regulates DRD1-mediated dopamine signal pathway. (A) N2a cells were transfected with VPS35-myc and control vectors for 24 h (left panels) or two mouse VPS35 shRNA (sh1 and sh2) and mock control shRNA vectors for 72 h (right panels). (B) N2a cells were transfected with DRD1 shRNA (shDRD1) and mock control shRNA vectors for 72 h. After equal splitting, cells were transfected with VPS35-myc and control vectors for 24 h. (C) N2a cells were transfected with mouse VPS35 shRNA (sh1 and sh2) and mock control shRNA vectors for 72 h. Cells were then treated with 50 μM dopamine (DOPA) or vehicle control (Ctrl) for another 36 h. After treatments, cells were lysed and equal protein amounts of cell lysates were subjected to Western blot analyses for indicated proteins. Protein levels were quantified by densitometry. Relative pCREB/CREB and pERK/ERK levels were calculated and compared to those of controls (set as one arbitrary units), n=3, * p
    Figure Legend Snippet: VPS35 regulates DRD1-mediated dopamine signal pathway. (A) N2a cells were transfected with VPS35-myc and control vectors for 24 h (left panels) or two mouse VPS35 shRNA (sh1 and sh2) and mock control shRNA vectors for 72 h (right panels). (B) N2a cells were transfected with DRD1 shRNA (shDRD1) and mock control shRNA vectors for 72 h. After equal splitting, cells were transfected with VPS35-myc and control vectors for 24 h. (C) N2a cells were transfected with mouse VPS35 shRNA (sh1 and sh2) and mock control shRNA vectors for 72 h. Cells were then treated with 50 μM dopamine (DOPA) or vehicle control (Ctrl) for another 36 h. After treatments, cells were lysed and equal protein amounts of cell lysates were subjected to Western blot analyses for indicated proteins. Protein levels were quantified by densitometry. Relative pCREB/CREB and pERK/ERK levels were calculated and compared to those of controls (set as one arbitrary units), n=3, * p

    Techniques Used: Transfection, shRNA, Western Blot

    21) Product Images from "Sulfuretin Attenuates MPP+-Induced Neurotoxicity through Akt/GSK3β and ERK Signaling Pathways"

    Article Title: Sulfuretin Attenuates MPP+-Induced Neurotoxicity through Akt/GSK3β and ERK Signaling Pathways

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms18122753

    LY294002 suppresses sulfuretin-induced protection against MPP + , whereas SB415286 reverses MPP + -induced cytotoxicity. SH-SY5Y cells were pretreated with or without LY294002 (10 μM) for 2 h, followed by treatment with or without sulfuretin (40 μM) for 2 h and exposed to MPP + (1 mM) for 24 h. ( A ) Cell viability was measured by MTT assay. Values are presented relative to control as mean percentage change ± S.D. ( n = 3). ( B ) Protein levels of p-Akt, Akt, p-GSK3β, GSK3β, p-ERK, ERK, and GAPDH were determined by Western blot analysis. Representative blots and their densitometric quantification are shown. Values are presented relative to control as mean fold change ± S.D. ( n = 3). ( C ) SH-SY5Y cells were pretreated with or without SB415286 (20 μM) for 2 h, and then exposed to MPP + (1 mM) for 24 h. Cell viability was measured by MTT assay. Values are presented relative to control as mean percentage change ± S.D. ( n = 3). Differences are statistically significant at ** p
    Figure Legend Snippet: LY294002 suppresses sulfuretin-induced protection against MPP + , whereas SB415286 reverses MPP + -induced cytotoxicity. SH-SY5Y cells were pretreated with or without LY294002 (10 μM) for 2 h, followed by treatment with or without sulfuretin (40 μM) for 2 h and exposed to MPP + (1 mM) for 24 h. ( A ) Cell viability was measured by MTT assay. Values are presented relative to control as mean percentage change ± S.D. ( n = 3). ( B ) Protein levels of p-Akt, Akt, p-GSK3β, GSK3β, p-ERK, ERK, and GAPDH were determined by Western blot analysis. Representative blots and their densitometric quantification are shown. Values are presented relative to control as mean fold change ± S.D. ( n = 3). ( C ) SH-SY5Y cells were pretreated with or without SB415286 (20 μM) for 2 h, and then exposed to MPP + (1 mM) for 24 h. Cell viability was measured by MTT assay. Values are presented relative to control as mean percentage change ± S.D. ( n = 3). Differences are statistically significant at ** p

    Techniques Used: MTT Assay, Western Blot

    MPP + decreases Akt/GSK3β and ERK phosphorylation and increases p53 expression, whereas sulfuretin reverses its effect. SH-SY5Y cells were pretreated with sulfuretin for 2 h and then treated with MPP + for 24 h. After cell lysis, the extracted proteins were subjected to Western blot analysis using specific antibodies. Protein levels of ( A ) p-Akt, Akt, p-GSK3β, GSK3β, p-CREB, and GAPDH; ( B ) p-ERK, ERK, p-p38, p38, p-JNK, and JNK were determined. Representative blots and their densitometric quantification are shown. Values are presented relative to control as mean fold change ± S.D. ( n = 3). Differences are statistically significant at * p
    Figure Legend Snippet: MPP + decreases Akt/GSK3β and ERK phosphorylation and increases p53 expression, whereas sulfuretin reverses its effect. SH-SY5Y cells were pretreated with sulfuretin for 2 h and then treated with MPP + for 24 h. After cell lysis, the extracted proteins were subjected to Western blot analysis using specific antibodies. Protein levels of ( A ) p-Akt, Akt, p-GSK3β, GSK3β, p-CREB, and GAPDH; ( B ) p-ERK, ERK, p-p38, p38, p-JNK, and JNK were determined. Representative blots and their densitometric quantification are shown. Values are presented relative to control as mean fold change ± S.D. ( n = 3). Differences are statistically significant at * p

    Techniques Used: Expressing, Lysis, Western Blot

    PD98059 suppresses sulfuretin-induced protection against MPP + . SH-SY5Y cells were pretreated with or without PD98059 (10 μM) for 2 h, followed by treatment with or without sulfuretin (40 μM) for 2 h, and exposed to MPP + (1 mM) for 24 h. ( A ) Cell viability was measured by MTT assay. Values are presented relative to control as mean percentage change ± S.D. ( n = 3). ( B ) Protein levels of p-ERK, ERK, p-Akt, Akt, p-GSK3β, GSK3β, and GAPDH were determined by Western blot analysis. Representative blots and their densitometric quantification are shown. Values are presented relative to control as mean fold change ± S.D. ( n = 3). Differences are statistically significant at * p
    Figure Legend Snippet: PD98059 suppresses sulfuretin-induced protection against MPP + . SH-SY5Y cells were pretreated with or without PD98059 (10 μM) for 2 h, followed by treatment with or without sulfuretin (40 μM) for 2 h, and exposed to MPP + (1 mM) for 24 h. ( A ) Cell viability was measured by MTT assay. Values are presented relative to control as mean percentage change ± S.D. ( n = 3). ( B ) Protein levels of p-ERK, ERK, p-Akt, Akt, p-GSK3β, GSK3β, and GAPDH were determined by Western blot analysis. Representative blots and their densitometric quantification are shown. Values are presented relative to control as mean fold change ± S.D. ( n = 3). Differences are statistically significant at * p

    Techniques Used: MTT Assay, Western Blot

    22) Product Images from "Effects of α-lipoic acid on LPS-induced neuroinflammation and NLRP3 inflammasome activation through the regulation of BV-2 microglial cells activation"

    Article Title: Effects of α-lipoic acid on LPS-induced neuroinflammation and NLRP3 inflammasome activation through the regulation of BV-2 microglial cells activation

    Journal: BMB Reports

    doi: 10.5483/BMBRep.2019.52.10.026

    Effects of α-LA on ERK, p38, NLRP3 inflammasome expression and NF-κB translocation in LPS-induced BV-2 microglial cells. BV-2 microglial cells were stimulated with LPS (1 μg/ml) for 30 min followed by treatment with the indicated concentrations of α-LA for 2 hours. (A) BV-2 microglial cells were lysed to whole lysates. Activation of ERK and p38 was detected by western blotting. (B) The translocation of NF-κB was also analyzed through western blotting. BV-2 microglial cells were lysed to cytosolic extracts and nucleic extracts, while both β-actin and Lamin-B1 were used as internal controls. (C) LPS and α-LA treatment was performed as described in the above legends. Finally, Iκ-Bα phosphorylation was detected by western blotting. (D) The western blot was performed to investigate the presence of NLRP3, pro-caspase-1 and cleaved caspase-1.
    Figure Legend Snippet: Effects of α-LA on ERK, p38, NLRP3 inflammasome expression and NF-κB translocation in LPS-induced BV-2 microglial cells. BV-2 microglial cells were stimulated with LPS (1 μg/ml) for 30 min followed by treatment with the indicated concentrations of α-LA for 2 hours. (A) BV-2 microglial cells were lysed to whole lysates. Activation of ERK and p38 was detected by western blotting. (B) The translocation of NF-κB was also analyzed through western blotting. BV-2 microglial cells were lysed to cytosolic extracts and nucleic extracts, while both β-actin and Lamin-B1 were used as internal controls. (C) LPS and α-LA treatment was performed as described in the above legends. Finally, Iκ-Bα phosphorylation was detected by western blotting. (D) The western blot was performed to investigate the presence of NLRP3, pro-caspase-1 and cleaved caspase-1.

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

    23) Product Images from "Inhibition of Stearoyl-CoA desaturase 1 reverts BRAF and MEK inhibition-induced selection of cancer stem cells in BRAF-mutated melanoma"

    Article Title: Inhibition of Stearoyl-CoA desaturase 1 reverts BRAF and MEK inhibition-induced selection of cancer stem cells in BRAF-mutated melanoma

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-018-0989-7

    SCD1 expression is able to predict the response of BRAF-mutated-melanoma cells to targeted agents. a - b ) AKT and ERK pathways were examined by WB analyses in protein lysates prepared from M14, A375 and M14-R cells treated with BRAF and MEK inhibitors or their combination (panel a ) grown in adhesion (2D) and as spheroids (3D) (panel b ); c ) SCD1 protein expression performed on fixed M14, A375 and M14-R spheroids after 96 h of treatment with BRAF and/or MEK inhibitors by Immunofluorescence analyses. Scale bar 50 μm; d ) WB analysis of SCD1 protein expression performed on M14-R and M14 spheroids after 96 h of BRAF and/or MEK inhibitors exposure; e ) Immunofluorescence analyses on YAP/TAZ expression were performed on fixed M14, Mel 29 and Mel 66 spheroids after 96 h of exposure to BRAF/MEK inhibitors; Scale bars: 10 μm; f ) Western blotting analysis of YAP/TAZ in M14, Mel 29 and Mel 66 spheroids after BRAF/MEK inhibitors exposure; g) YAP/TAZ downstream target ctgf , cyr61 , birc5 and tead4 expression in A375, M14, Mel 29 and Mel 66 by qRT-PCR analyses
    Figure Legend Snippet: SCD1 expression is able to predict the response of BRAF-mutated-melanoma cells to targeted agents. a - b ) AKT and ERK pathways were examined by WB analyses in protein lysates prepared from M14, A375 and M14-R cells treated with BRAF and MEK inhibitors or their combination (panel a ) grown in adhesion (2D) and as spheroids (3D) (panel b ); c ) SCD1 protein expression performed on fixed M14, A375 and M14-R spheroids after 96 h of treatment with BRAF and/or MEK inhibitors by Immunofluorescence analyses. Scale bar 50 μm; d ) WB analysis of SCD1 protein expression performed on M14-R and M14 spheroids after 96 h of BRAF and/or MEK inhibitors exposure; e ) Immunofluorescence analyses on YAP/TAZ expression were performed on fixed M14, Mel 29 and Mel 66 spheroids after 96 h of exposure to BRAF/MEK inhibitors; Scale bars: 10 μm; f ) Western blotting analysis of YAP/TAZ in M14, Mel 29 and Mel 66 spheroids after BRAF/MEK inhibitors exposure; g) YAP/TAZ downstream target ctgf , cyr61 , birc5 and tead4 expression in A375, M14, Mel 29 and Mel 66 by qRT-PCR analyses

    Techniques Used: Expressing, Western Blot, Immunofluorescence, Quantitative RT-PCR

    24) Product Images from "P2Y6 contributes to ovalbumin-induced allergic asthma by enhancing mast cell function in mice"

    Article Title: P2Y6 contributes to ovalbumin-induced allergic asthma by enhancing mast cell function in mice

    Journal: Oncotarget

    doi: 10.18632/oncotarget.11758

    P2Y 6 -related signaling pathway detection by western blot in mast cells A. The phosphorylation levels of P65, AKT, ERK, and P38 in mast cells were determined by western blot. B. The phosphorylation levels of AKT and ERK in the lung tissues in ovalbumin-induced wild type and P2Y −/− mice. C. The phosphorylation levels of AKT and ERK in the lung tissues in UDP-treated wild type and P2Y −/− asthmatic mice. WT is the abbreviation of wild type; UDP is the abbreviation of uridine 5′-diphosphate; OVA is the abbreviation of ovalbumin.
    Figure Legend Snippet: P2Y 6 -related signaling pathway detection by western blot in mast cells A. The phosphorylation levels of P65, AKT, ERK, and P38 in mast cells were determined by western blot. B. The phosphorylation levels of AKT and ERK in the lung tissues in ovalbumin-induced wild type and P2Y −/− mice. C. The phosphorylation levels of AKT and ERK in the lung tissues in UDP-treated wild type and P2Y −/− asthmatic mice. WT is the abbreviation of wild type; UDP is the abbreviation of uridine 5′-diphosphate; OVA is the abbreviation of ovalbumin.

    Techniques Used: Western Blot, Mouse Assay

    25) Product Images from "Exosomal microRNA miR-1246 induces cell motility and invasion through the regulation of DENND2D in oral squamous cell carcinoma"

    Article Title: Exosomal microRNA miR-1246 induces cell motility and invasion through the regulation of DENND2D in oral squamous cell carcinoma

    Journal: Scientific Reports

    doi: 10.1038/srep38750

    LM-exosomes increase cell viability and cell motility in HOC313-P cells. ( a ) HOC313-P cells were treated with PBS or LM-exosomes, and the effect of exosomes on cell growth was determined using an in vitro WST-8 assay at the indicated times. ( b ) Western blot analysis of EGFR, ERK and AKT and their phosphorylation status is shown in HOC313-P cells treated with LM-exosomes for the indicated amounts of time. ( c and d ) Cell migration was assessed by Transwell migration assay ( c ) and cell invasion was assessed by Transwell invasion assay ( d ) in HOC313-P cells treated with either PBS or LM-exosomes. Experiments were performed in triplicate. (Bars, SD). Student’s t -test was used for statistical analysis; asterisks represent P
    Figure Legend Snippet: LM-exosomes increase cell viability and cell motility in HOC313-P cells. ( a ) HOC313-P cells were treated with PBS or LM-exosomes, and the effect of exosomes on cell growth was determined using an in vitro WST-8 assay at the indicated times. ( b ) Western blot analysis of EGFR, ERK and AKT and their phosphorylation status is shown in HOC313-P cells treated with LM-exosomes for the indicated amounts of time. ( c and d ) Cell migration was assessed by Transwell migration assay ( c ) and cell invasion was assessed by Transwell invasion assay ( d ) in HOC313-P cells treated with either PBS or LM-exosomes. Experiments were performed in triplicate. (Bars, SD). Student’s t -test was used for statistical analysis; asterisks represent P

    Techniques Used: In Vitro, Western Blot, Migration, Transwell Migration Assay, Transwell Invasion Assay

    26) Product Images from "DEC2 suppresses tumor proliferation and metastasis by regulating ERK/NF-κB pathway in gastric cancer"

    Article Title: DEC2 suppresses tumor proliferation and metastasis by regulating ERK/NF-κB pathway in gastric cancer

    Journal: American Journal of Cancer Research

    doi:

    . B, E. Western blot results of NF-κB p65, p-ERK, and E-cadherin expression levels in DEC2/MGC803, DEC2/MKN-45, DEC2-sh/HGC27 and their corresponding control cells after treated with ERK inhibitor U0126 (10 μM). C. Comparisons of the migration and invasion ability in stable transfected DEC2/MGC803 and DEC2/MKN-45 cells after treated with ERK inhibitor U0126 (10 μM) or not. Quantitative results are shown in the right panel. F. Comparisons of the migration and invasion ability in DEC2-sh/HGC27 and their corresponding control cells after treated with ERK inhibitor U0126 (10 μM) or not. Quantitative results are shown in the bottom panel. The error bars indicate ± SEM. *P
    Figure Legend Snippet: . B, E. Western blot results of NF-κB p65, p-ERK, and E-cadherin expression levels in DEC2/MGC803, DEC2/MKN-45, DEC2-sh/HGC27 and their corresponding control cells after treated with ERK inhibitor U0126 (10 μM). C. Comparisons of the migration and invasion ability in stable transfected DEC2/MGC803 and DEC2/MKN-45 cells after treated with ERK inhibitor U0126 (10 μM) or not. Quantitative results are shown in the right panel. F. Comparisons of the migration and invasion ability in DEC2-sh/HGC27 and their corresponding control cells after treated with ERK inhibitor U0126 (10 μM) or not. Quantitative results are shown in the bottom panel. The error bars indicate ± SEM. *P

    Techniques Used: Western Blot, Expressing, Migration, Transfection

    27) Product Images from "Hydrogen peroxide mediates hyperglycemia-induced invasive activity via ERK and p38 MAPK in human pancreatic cancer"

    Article Title: Hydrogen peroxide mediates hyperglycemia-induced invasive activity via ERK and p38 MAPK in human pancreatic cancer

    Journal: Oncotarget

    doi:

    HG activates MAPK pathways and the NF-κB and AP-1 transcription factors via the production of H 2 O 2 in BxPC-3 and Panc-1 cells A. The effect of 25 mM glucose on the phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun. PC cells were treated with 25 mM glucose for the indicated times. Phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun were determined using Western blot analysis. B. The effect of H 2 O 2 (200 μM) on the phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun. PC cells were treated with 200 μM H 2 O 2 for the indicated times. Phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun were determined using Western blot analysis. C. The effect of MAPK pathway inhibitors on the phosphorylation of ERK and p38. D. The effect of MAPK pathway inhibitors on the phosphorylation of NF-κB and c-Jun. BxPC-3 and Panc-1 cells were treated with the selective MAPK pathway inhibitors PD 98059 (50 μM) and SB 203580 (20 μM), as well as PEG-CAT (1000 U/ml) in the presence or absence of high glucose concentrations. The phosphorylation of ERK, p38 (C), NF-κB and c-Jun (D) were analyzed using Western blot analysis for 24 h. E. The effect of SOD2 knockdown on the phosphorylation of ERK and p38 in the presence or absence of high glucose concentrations. F. The effect of SOD2 knockdown on the phosphorylation of NF-κB and c-Jun in the presence or absence of high glucose concentrations. After the BxPC-3 and Panc-1 cells were transfected with siRNAs for 48 h, the phosphorylation levels of ERK, p38 (E), NF-κB and c-Jun (F) were determined using Western blot analysis.
    Figure Legend Snippet: HG activates MAPK pathways and the NF-κB and AP-1 transcription factors via the production of H 2 O 2 in BxPC-3 and Panc-1 cells A. The effect of 25 mM glucose on the phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun. PC cells were treated with 25 mM glucose for the indicated times. Phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun were determined using Western blot analysis. B. The effect of H 2 O 2 (200 μM) on the phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun. PC cells were treated with 200 μM H 2 O 2 for the indicated times. Phosphorylation of ERK, p38 MAPK, NF-κB and c-Jun were determined using Western blot analysis. C. The effect of MAPK pathway inhibitors on the phosphorylation of ERK and p38. D. The effect of MAPK pathway inhibitors on the phosphorylation of NF-κB and c-Jun. BxPC-3 and Panc-1 cells were treated with the selective MAPK pathway inhibitors PD 98059 (50 μM) and SB 203580 (20 μM), as well as PEG-CAT (1000 U/ml) in the presence or absence of high glucose concentrations. The phosphorylation of ERK, p38 (C), NF-κB and c-Jun (D) were analyzed using Western blot analysis for 24 h. E. The effect of SOD2 knockdown on the phosphorylation of ERK and p38 in the presence or absence of high glucose concentrations. F. The effect of SOD2 knockdown on the phosphorylation of NF-κB and c-Jun in the presence or absence of high glucose concentrations. After the BxPC-3 and Panc-1 cells were transfected with siRNAs for 48 h, the phosphorylation levels of ERK, p38 (E), NF-κB and c-Jun (F) were determined using Western blot analysis.

    Techniques Used: Western Blot, Transfection

    28) Product Images from "A SQSTM1/p62 mutation linked to Paget's disease increases the osteoclastogenic potential of the bone microenvironment"

    Article Title: A SQSTM1/p62 mutation linked to Paget's disease increases the osteoclastogenic potential of the bone microenvironment

    Journal:

    doi: 10.1093/hmg/ddn266

    Activation of the p38 MAPK and ERK 1/2 signaling pathways in KI or WT stromal cells induced by 1α,25-(OH) 2 D 3 : stromal cells from KI or WT mice were cultured with IMDM + 10% FCS for 14 days. Cells were then cultured with IMDM + 2% FCS for 24 h
    Figure Legend Snippet: Activation of the p38 MAPK and ERK 1/2 signaling pathways in KI or WT stromal cells induced by 1α,25-(OH) 2 D 3 : stromal cells from KI or WT mice were cultured with IMDM + 10% FCS for 14 days. Cells were then cultured with IMDM + 2% FCS for 24 h

    Techniques Used: Activation Assay, Mouse Assay, Cell Culture

    29) Product Images from "Enzymatic cleavage of myoferlin releases a dual C2-domain module linked to ERK signalling"

    Article Title: Enzymatic cleavage of myoferlin releases a dual C2-domain module linked to ERK signalling

    Journal: Cellular signalling

    doi: 10.1016/j.cellsig.2017.02.009

    Overexpression of canonical full-length myoferlin (MFL) but not the uncleavable M38a, upregulates phosphorylated ERK in transfected HEK293 cells. (A) Cell lysates from transfected HEK293 cells were incubated with Proteome Profiler™ Antibody Arrays (Human Phospho-MAPK and Mouse-RTK Array Kit, R D systems). Histogram of densitometry from duplicate spots of 13 representative proteins from the Human Phospho-MAPK Array showing elevation of phospho-ERK. No differences between M FL and M 38a or empty pIRES were observed for the remaining phosphoproteins (not shown). (B and C) Western blot experiments confirm specific upregulation of pERK relative to total ERK in HEK293 cells transfected with M FL , but not M 38a , D 40a or empty pIRES. Levels of pAKT were unchanged. Data show three experiments performed in triplicate. (D) Densitometric analysis of combined data from western blots showing up-regulation of pERK in M FL expressing HEK293 cells was significantly elevated relative to pIRES expressing cells ( p = 0.0002) (one way ANOVA, Prism 6).
    Figure Legend Snippet: Overexpression of canonical full-length myoferlin (MFL) but not the uncleavable M38a, upregulates phosphorylated ERK in transfected HEK293 cells. (A) Cell lysates from transfected HEK293 cells were incubated with Proteome Profiler™ Antibody Arrays (Human Phospho-MAPK and Mouse-RTK Array Kit, R D systems). Histogram of densitometry from duplicate spots of 13 representative proteins from the Human Phospho-MAPK Array showing elevation of phospho-ERK. No differences between M FL and M 38a or empty pIRES were observed for the remaining phosphoproteins (not shown). (B and C) Western blot experiments confirm specific upregulation of pERK relative to total ERK in HEK293 cells transfected with M FL , but not M 38a , D 40a or empty pIRES. Levels of pAKT were unchanged. Data show three experiments performed in triplicate. (D) Densitometric analysis of combined data from western blots showing up-regulation of pERK in M FL expressing HEK293 cells was significantly elevated relative to pIRES expressing cells ( p = 0.0002) (one way ANOVA, Prism 6).

    Techniques Used: Over Expression, Transfection, Incubation, Western Blot, Expressing

    30) Product Images from "Trichomonas vaginalis Induces Production of Proinflammatory Cytokines in Mouse Macrophages Through Activation of MAPK and NF-κB Pathways Partially Mediated by TLR2"

    Article Title: Trichomonas vaginalis Induces Production of Proinflammatory Cytokines in Mouse Macrophages Through Activation of MAPK and NF-κB Pathways Partially Mediated by TLR2

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00712

    Trichomonas vaginalis activates p38 and ERK signal pathways in WT mouse peritoneal macrophages. WT macrophages were co-incubated with T. vaginalis for different times (0, 0.5, 1, 2, and 4 h), phosphorylation of p38 and ERK were detected by western blot (A) . Relative Gray analysis of western blot (B) . The phosphorylated of p38 (at 0.5 h) and ERK (at 2 h) were observed obviously. Data are expressed as the mean ± SD from three separate experiments ( ∗ p
    Figure Legend Snippet: Trichomonas vaginalis activates p38 and ERK signal pathways in WT mouse peritoneal macrophages. WT macrophages were co-incubated with T. vaginalis for different times (0, 0.5, 1, 2, and 4 h), phosphorylation of p38 and ERK were detected by western blot (A) . Relative Gray analysis of western blot (B) . The phosphorylated of p38 (at 0.5 h) and ERK (at 2 h) were observed obviously. Data are expressed as the mean ± SD from three separate experiments ( ∗ p

    Techniques Used: Incubation, Western Blot

    Trichomonas vaginalis induces cytokines production regulated by p38 and ERK via TLR2. Phosphorylationof p38 and ERK in TLR2 -/- mouse peritoneal macrophages were significantly reduced after co-incubated with T. vaginalis for 0.5 h (p38) or 2 h (ERK) compared to in WT mouse peritoneal macrophages (A) . Inhibitors of p38 (SB203580; 30 μM) or ERK (PD98059; 40 μM) were used to pretreated WT mouse peritoneal macrophages for 1 h before co-incubated by T. vaginalis . Phosphorylation of p38 and ERK in SB203580 and PD98059 pre-treated WT macrophages (C) . Relative Gray analysis of western blot (B,D) . The production of IL-6, TNF-α, and IFN-γ induced by T. vaginalis were significantly inhibited by the inhibitors compared to in WT mouse peritoneal macrophages (E–G) . Data are expressed as the mean ± SD from three separate experiments ( ∗ p
    Figure Legend Snippet: Trichomonas vaginalis induces cytokines production regulated by p38 and ERK via TLR2. Phosphorylationof p38 and ERK in TLR2 -/- mouse peritoneal macrophages were significantly reduced after co-incubated with T. vaginalis for 0.5 h (p38) or 2 h (ERK) compared to in WT mouse peritoneal macrophages (A) . Inhibitors of p38 (SB203580; 30 μM) or ERK (PD98059; 40 μM) were used to pretreated WT mouse peritoneal macrophages for 1 h before co-incubated by T. vaginalis . Phosphorylation of p38 and ERK in SB203580 and PD98059 pre-treated WT macrophages (C) . Relative Gray analysis of western blot (B,D) . The production of IL-6, TNF-α, and IFN-γ induced by T. vaginalis were significantly inhibited by the inhibitors compared to in WT mouse peritoneal macrophages (E–G) . Data are expressed as the mean ± SD from three separate experiments ( ∗ p

    Techniques Used: Incubation, Western Blot

    31) Product Images from "KCa3.1 mediates activation of fibroblasts in diabetic renal interstitial fibrosis"

    Article Title: KCa3.1 mediates activation of fibroblasts in diabetic renal interstitial fibrosis

    Journal: Nephrology Dialysis Transplantation

    doi: 10.1093/ndt/gft431

    KCa3.1 mediated TGF-β1-induced activation of renal cortical fibroblasts and fibrotic responses through Smad and ERK pathway but not P38 and JNK pathways. Human renal interstitial fibroblasts were treated with control, TGF-β1 (2 ng/mL) or TGF-β1 (2 ng/mL) combined with TRAM34 (2 μM) for 48 h. Western blot results showed that TRAM34 inhibited the TGF-β1-mediated increases in p-Smad2 ( A ), p-Smad3 ( B ) expression and p-ERK1/2 expression ( C ). However, TRAM34 had no effect on the P38 ( D ) and JNK ( E ) MAP kinase pathways. Results are presented as means ± SEM. *P
    Figure Legend Snippet: KCa3.1 mediated TGF-β1-induced activation of renal cortical fibroblasts and fibrotic responses through Smad and ERK pathway but not P38 and JNK pathways. Human renal interstitial fibroblasts were treated with control, TGF-β1 (2 ng/mL) or TGF-β1 (2 ng/mL) combined with TRAM34 (2 μM) for 48 h. Western blot results showed that TRAM34 inhibited the TGF-β1-mediated increases in p-Smad2 ( A ), p-Smad3 ( B ) expression and p-ERK1/2 expression ( C ). However, TRAM34 had no effect on the P38 ( D ) and JNK ( E ) MAP kinase pathways. Results are presented as means ± SEM. *P

    Techniques Used: Activation Assay, Western Blot, Expressing

    32) Product Images from "CD73/NT5E is a target of miR-30a-5p and plays an important role in the pathogenesis of non-small cell lung cancer"

    Article Title: CD73/NT5E is a target of miR-30a-5p and plays an important role in the pathogenesis of non-small cell lung cancer

    Journal: Molecular Cancer

    doi: 10.1186/s12943-017-0591-1

    Overexpression of miR-30a-5p inhibits NSCLC cell proliferation and motility. a b CCK-8 assay of cell viability in NSCLC cell lines transfected with miR-30a-5p mimics or miR-30a-5p inhibitor at 24, 48 and 72 h. c A wound healing assay was performed to observe the role of miR-30a-5p transfection in cells. The data showed that the speed with which the cells migrated towards the scratch was lower in cells transfected with the miR-30a-5p overexpression vector than in the control cells. d Overexpression of miR-30a-5p inhibits invasion and migration of NSCLC cells. The A549 and H226 cell lines were transfected with miR-30a-5p mimics and allowed to migrate through 8-μM pore Transwells. The cells that migrated were stained and counted in at least three microscopic fields (magnification, ×100). Then, cells were treated as described before and allowed to invade through the Matrigel-coated membrane in Transwells. The invasive cells were stained and counted under a light microscope. e and f The A549 and H226 cells were treated with or without miR-30a-5p mimics or the miR-30a-5p inhibitor for 72 h, respectively. The expression levels of p-EGFR, EGFR, p-AKT, AKT, p-ERK, ERK and Cyclin D1 were analyzed by western blotting. ** P
    Figure Legend Snippet: Overexpression of miR-30a-5p inhibits NSCLC cell proliferation and motility. a b CCK-8 assay of cell viability in NSCLC cell lines transfected with miR-30a-5p mimics or miR-30a-5p inhibitor at 24, 48 and 72 h. c A wound healing assay was performed to observe the role of miR-30a-5p transfection in cells. The data showed that the speed with which the cells migrated towards the scratch was lower in cells transfected with the miR-30a-5p overexpression vector than in the control cells. d Overexpression of miR-30a-5p inhibits invasion and migration of NSCLC cells. The A549 and H226 cell lines were transfected with miR-30a-5p mimics and allowed to migrate through 8-μM pore Transwells. The cells that migrated were stained and counted in at least three microscopic fields (magnification, ×100). Then, cells were treated as described before and allowed to invade through the Matrigel-coated membrane in Transwells. The invasive cells were stained and counted under a light microscope. e and f The A549 and H226 cells were treated with or without miR-30a-5p mimics or the miR-30a-5p inhibitor for 72 h, respectively. The expression levels of p-EGFR, EGFR, p-AKT, AKT, p-ERK, ERK and Cyclin D1 were analyzed by western blotting. ** P

    Techniques Used: Over Expression, CCK-8 Assay, Transfection, Wound Healing Assay, Plasmid Preparation, Migration, Staining, Light Microscopy, Expressing, Western Blot

    CD73 knockdown increased the sensitivity of NSCLC cells to gefitinib and lapatinib. a The expression of proteins in the CD73 and EGFR signaling pathways was detected in the stable A549 and H226 cell lines. Our data showed that p-EGFR and p-AKT expression is significantly decreased in CD73-silenced cell lines compared with the control cells. b After serum starvation for 24 h, CD73-silenced A549 and H226 cells were treated with or without EGF (50 ng/ml) for 30 min. The expression levels of CD73, p-EGFR, EGFR, p-AKT, AKT, p-ERK and ERK were analyzed by western blot analysis. c – f CD73 knockdown increased the sensitivity of NSCLC cells to gefitinib and lapatinib. The A549 and H226 cell lines were transfected with 10 μM gefitinib and 20 μM lapatinib for 48 h, respectively. After the aforementioned treatments, cell viability was assessed using CCK-8 assays. The data shown represent the mean ± SD values of four replicate experiments. All the data were obtained from three independent experiments and are shown as the mean ± SD values. ** P 0.01; *** P
    Figure Legend Snippet: CD73 knockdown increased the sensitivity of NSCLC cells to gefitinib and lapatinib. a The expression of proteins in the CD73 and EGFR signaling pathways was detected in the stable A549 and H226 cell lines. Our data showed that p-EGFR and p-AKT expression is significantly decreased in CD73-silenced cell lines compared with the control cells. b After serum starvation for 24 h, CD73-silenced A549 and H226 cells were treated with or without EGF (50 ng/ml) for 30 min. The expression levels of CD73, p-EGFR, EGFR, p-AKT, AKT, p-ERK and ERK were analyzed by western blot analysis. c – f CD73 knockdown increased the sensitivity of NSCLC cells to gefitinib and lapatinib. The A549 and H226 cell lines were transfected with 10 μM gefitinib and 20 μM lapatinib for 48 h, respectively. After the aforementioned treatments, cell viability was assessed using CCK-8 assays. The data shown represent the mean ± SD values of four replicate experiments. All the data were obtained from three independent experiments and are shown as the mean ± SD values. ** P 0.01; *** P

    Techniques Used: Expressing, Western Blot, Transfection, CCK-8 Assay

    33) Product Images from "Critical roles for Nitric oxide and ERK in the completion of prosurvival autophagy in 4OHTAM-treated estrogen receptor-positive breast cancer cells"

    Article Title: Critical roles for Nitric oxide and ERK in the completion of prosurvival autophagy in 4OHTAM-treated estrogen receptor-positive breast cancer cells

    Journal: Cancer letters

    doi: 10.1016/j.canlet.2014.07.031

    O2- mediates 4OHTAM-induced activation of ERK which is enhanced by c-PTIO critical for lysosomal stability and survival in MCF7 cells The cells were treated with the indicated drugs (5 μM of 4OHTAM, 100 μM c-PTIO, 2 mM of Tiron and 20 μM of PD98059) for two days. Whole cell lysates were immunoblotted for phosphorylated and total protein of ERK, JNK, and p38 MAPK (A) and ERK and LC3 (B). C. Cells treated with 4OHTAM (5 μM) and PD98059 (20 μM) and analyzed for MDC sequestration. D. The cells were treated with the indicated drugs for 3 days, labeled with AO and measured the cellular fluorescence. Relative AO red/green ratio was presented as a graph with standard deviation indicated. E. The cells were treated for 3 days, labeled with AO and measured at the indicated time points, and the average red/green ratio normalized to the non-treatment condition was presented with standard deviation indicated. F. Cell viability was analyzed by MTT assay. There are significant differences between 4OHTAM and combination of 4OHTAM with c-PTIO or PD98059 (p
    Figure Legend Snippet: O2- mediates 4OHTAM-induced activation of ERK which is enhanced by c-PTIO critical for lysosomal stability and survival in MCF7 cells The cells were treated with the indicated drugs (5 μM of 4OHTAM, 100 μM c-PTIO, 2 mM of Tiron and 20 μM of PD98059) for two days. Whole cell lysates were immunoblotted for phosphorylated and total protein of ERK, JNK, and p38 MAPK (A) and ERK and LC3 (B). C. Cells treated with 4OHTAM (5 μM) and PD98059 (20 μM) and analyzed for MDC sequestration. D. The cells were treated with the indicated drugs for 3 days, labeled with AO and measured the cellular fluorescence. Relative AO red/green ratio was presented as a graph with standard deviation indicated. E. The cells were treated for 3 days, labeled with AO and measured at the indicated time points, and the average red/green ratio normalized to the non-treatment condition was presented with standard deviation indicated. F. Cell viability was analyzed by MTT assay. There are significant differences between 4OHTAM and combination of 4OHTAM with c-PTIO or PD98059 (p

    Techniques Used: Activation Assay, Labeling, Fluorescence, Standard Deviation, MTT Assay

    34) Product Images from "Regulation of the Maintenance of Peripheral T-Cell Anergy by TAB1-Mediated p38? Activation"

    Article Title: Regulation of the Maintenance of Peripheral T-Cell Anergy by TAB1-Mediated p38? Activation

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.24.16.6957-6966.2004

    Anergic CD4 + T cells show decreased activation of ERK and JNK and increased activation of p38 MAPK in response to TCR stimulation. (A) CD4 + T cells were purified from untreated Vβ8.1-tg mice (naive) or Mls-1 a -treated Vβ8.1-tg mice (anergic) by treatment of lymph node cells with anti-CD8 MAb and complement, followed by nylon wool column enrichment. Cells were cultured for 3 h at 37°C in complete medium to decrease basal levels of MAPK activity. Cells then were placed in anti-TCR MAb (10 μg/ml)-coated plates for various times and lysed, and the levels of phosphorylated and total MAPKs were determined by immunoblotting with specific antibodies. Representative results from three independent experiments are shown. (B) In vitro immune complex kinase assay of p38α and ERK1. Naive or anergic CD4 + T cells were prepared as described for panel A and placed in anti-TCR MAb (10 μg/ml)-coated plates in the presence or absence of SB203580 for various times. Cell lysates were immunoprecipitated with anti-p38α or anti-ERK1 antibody, and in vitro kinase assays of the immunoprecipitates were performed with ATF2 and maltose-binding protein (MBP) as substrates for p38α and ERK1, respectively. Proteins were separated by SDS-PAGE and analyzed by autoradiography. The quantities of p38α and ERK1 were assessed by probing the immunoblots with antibodies specific for p38α or ERK1. Representative results from three independent experiments are shown.
    Figure Legend Snippet: Anergic CD4 + T cells show decreased activation of ERK and JNK and increased activation of p38 MAPK in response to TCR stimulation. (A) CD4 + T cells were purified from untreated Vβ8.1-tg mice (naive) or Mls-1 a -treated Vβ8.1-tg mice (anergic) by treatment of lymph node cells with anti-CD8 MAb and complement, followed by nylon wool column enrichment. Cells were cultured for 3 h at 37°C in complete medium to decrease basal levels of MAPK activity. Cells then were placed in anti-TCR MAb (10 μg/ml)-coated plates for various times and lysed, and the levels of phosphorylated and total MAPKs were determined by immunoblotting with specific antibodies. Representative results from three independent experiments are shown. (B) In vitro immune complex kinase assay of p38α and ERK1. Naive or anergic CD4 + T cells were prepared as described for panel A and placed in anti-TCR MAb (10 μg/ml)-coated plates in the presence or absence of SB203580 for various times. Cell lysates were immunoprecipitated with anti-p38α or anti-ERK1 antibody, and in vitro kinase assays of the immunoprecipitates were performed with ATF2 and maltose-binding protein (MBP) as substrates for p38α and ERK1, respectively. Proteins were separated by SDS-PAGE and analyzed by autoradiography. The quantities of p38α and ERK1 were assessed by probing the immunoblots with antibodies specific for p38α or ERK1. Representative results from three independent experiments are shown.

    Techniques Used: Activation Assay, Purification, Mouse Assay, Cell Culture, Activity Assay, In Vitro, Immune Complex Kinase Assay, Immunoprecipitation, Binding Assay, SDS Page, Autoradiography, Western Blot

    The inhibition of ERK kinase activity by TAB1 is dependent on p38. (A) Mock-2B4 cells (Mock) and TAB1-2B4 cells (TAB1) were cultured with various doses of PMA (0 to 60 ng/ml) for 10 min, and the levels of phosphorylated ERK and total ERK were determined by immunoblotting with specific antibodies. (B) Mock-2B4 cells (Mock) and TAB1-2B4 cells (TAB1) were cultured in the presence (SB) or absence (Cont) of SB203580 (10 μM) for 2 h and subsequently stimulated with PMA (0, 10, or 20 ng/ml) for 10 min. Cells were lysed, and the levels of phosphorylated ERK and total ERK were determined by immunoblotting with specific antibodies. (C) Mock-2B4 cells were transduced with a retroviral supernatant containing empty vector (Mock/Mock) or p38DN (Mock/p38DN) to establish stable cell lines. TAB1-2B4 cells were also transduced to establish cell lines expressing empty vector (TAB1/Mock) or p38DN cDNA (TAB1/p38DN). These cell lines were stimulated with PMA (0, 5, or 25 ng/ml) for 10 min and lysed, and the levels of phosphorylated MAPks and total MAPKs were determined by immunoblotting with specific antibodies. Representative results from two independent experiments are shown. (D) Mock-2B4 cells (Mock) and TAB1-2B4 cells (TAB1) were treated or not treated with SB203580 (10 μM) for 2 h and stimulated with PMA (20 ng/ml) and ionomycin (Iono; 2 μM) for 8 h in the presence (SB) or absence (Cont) of SB203580, respectively. Cells were subjected to RT-PCR analysis for IL-2 and G3PDH mRNAs. Representative results from two independent experiments are shown. (E) Mock/Mock, Mock/p38DN, TAB1/Mock, and TAB1/p38DN cells were stimulated with PMA (0, 5, or 25 ng/ml) and ionomycin (2 μM) for 8 h. Cells were subjected to RT-PCR analysis for IL-2, IL-10, and G3PDH mRNAs. Representative results from two independent experiments are shown. (F) Mock/Mock, Mock/p38DN, TAB1/Mock, and TAB1/p38DN cells were stimulated with anti-TCR MAb (H57; 10 μg/ml) for 24 h. The levels of IL-2 in culture supernatants were determined by an ELISA. IL-2 was not detectable in cultures without anti-TCR MAb.
    Figure Legend Snippet: The inhibition of ERK kinase activity by TAB1 is dependent on p38. (A) Mock-2B4 cells (Mock) and TAB1-2B4 cells (TAB1) were cultured with various doses of PMA (0 to 60 ng/ml) for 10 min, and the levels of phosphorylated ERK and total ERK were determined by immunoblotting with specific antibodies. (B) Mock-2B4 cells (Mock) and TAB1-2B4 cells (TAB1) were cultured in the presence (SB) or absence (Cont) of SB203580 (10 μM) for 2 h and subsequently stimulated with PMA (0, 10, or 20 ng/ml) for 10 min. Cells were lysed, and the levels of phosphorylated ERK and total ERK were determined by immunoblotting with specific antibodies. (C) Mock-2B4 cells were transduced with a retroviral supernatant containing empty vector (Mock/Mock) or p38DN (Mock/p38DN) to establish stable cell lines. TAB1-2B4 cells were also transduced to establish cell lines expressing empty vector (TAB1/Mock) or p38DN cDNA (TAB1/p38DN). These cell lines were stimulated with PMA (0, 5, or 25 ng/ml) for 10 min and lysed, and the levels of phosphorylated MAPks and total MAPKs were determined by immunoblotting with specific antibodies. Representative results from two independent experiments are shown. (D) Mock-2B4 cells (Mock) and TAB1-2B4 cells (TAB1) were treated or not treated with SB203580 (10 μM) for 2 h and stimulated with PMA (20 ng/ml) and ionomycin (Iono; 2 μM) for 8 h in the presence (SB) or absence (Cont) of SB203580, respectively. Cells were subjected to RT-PCR analysis for IL-2 and G3PDH mRNAs. Representative results from two independent experiments are shown. (E) Mock/Mock, Mock/p38DN, TAB1/Mock, and TAB1/p38DN cells were stimulated with PMA (0, 5, or 25 ng/ml) and ionomycin (2 μM) for 8 h. Cells were subjected to RT-PCR analysis for IL-2, IL-10, and G3PDH mRNAs. Representative results from two independent experiments are shown. (F) Mock/Mock, Mock/p38DN, TAB1/Mock, and TAB1/p38DN cells were stimulated with anti-TCR MAb (H57; 10 μg/ml) for 24 h. The levels of IL-2 in culture supernatants were determined by an ELISA. IL-2 was not detectable in cultures without anti-TCR MAb.

    Techniques Used: Inhibition, Activity Assay, Cell Culture, Transduction, Plasmid Preparation, Stable Transfection, Expressing, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    35) Product Images from "Plant Polyphenols Regulate Chemokine Expression and Tissue Repair in Human Keratinocytes Through Interaction with Cytoplasmic and Nuclear Components of Epidermal Growth Factor Receptor System"

    Article Title: Plant Polyphenols Regulate Chemokine Expression and Tissue Repair in Human Keratinocytes Through Interaction with Cytoplasmic and Nuclear Components of Epidermal Growth Factor Receptor System

    Journal: Antioxidants & Redox Signaling

    doi: 10.1089/ars.2011.4053

    PP effects on (tumor necrosis factor alpha [TNF-α] plus IFN-γ ) -induced EGFR, ERK, and p65 phosphorylation, chemokine production, and nuclear factor κB (NFκB) and activator protein 1 (AP-1) DNA binding. (A) Concentration-dependent
    Figure Legend Snippet: PP effects on (tumor necrosis factor alpha [TNF-α] plus IFN-γ ) -induced EGFR, ERK, and p65 phosphorylation, chemokine production, and nuclear factor κB (NFκB) and activator protein 1 (AP-1) DNA binding. (A) Concentration-dependent

    Techniques Used: Binding Assay, Concentration Assay

    PP effects on spontaneous epidermal growth factor receptor (EGFR)/extracellular regulation kinase (ERK)/p65 phosphorylation, chemokine gene expression, and protein synthesis in normal human epidermal keratinocytes (NHEK). (A) Western blots of EGFR and
    Figure Legend Snippet: PP effects on spontaneous epidermal growth factor receptor (EGFR)/extracellular regulation kinase (ERK)/p65 phosphorylation, chemokine gene expression, and protein synthesis in normal human epidermal keratinocytes (NHEK). (A) Western blots of EGFR and

    Techniques Used: Expressing, Western Blot

    36) Product Images from "JDP2 and ATF3 deficiencies dampen maladaptive cardiac remodeling and preserve cardiac function"

    Article Title: JDP2 and ATF3 deficiencies dampen maladaptive cardiac remodeling and preserve cardiac function

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0213081

    dKO male mice display increased p38 activity. Cardiac hypertrophy was induced by TAC in male mice. Eight weeks following TAC, mice were sacrificed and hearts were excised A Western blot analysis of heart lysate derived from the indicated genotypes with the indicated antibodies. B-C Densitometric analysis of Western blot shown in A is presented as mean ratio of the corresponding phospho-protein to total protein ± SE (compared to wild type control, defined as 1, n = 5-6/group). B pp38/p38. C pERK/ERK. *** P ≤ 0.05, control vs. TAC; † P ≤ 0.05 difference between genotypes.
    Figure Legend Snippet: dKO male mice display increased p38 activity. Cardiac hypertrophy was induced by TAC in male mice. Eight weeks following TAC, mice were sacrificed and hearts were excised A Western blot analysis of heart lysate derived from the indicated genotypes with the indicated antibodies. B-C Densitometric analysis of Western blot shown in A is presented as mean ratio of the corresponding phospho-protein to total protein ± SE (compared to wild type control, defined as 1, n = 5-6/group). B pp38/p38. C pERK/ERK. *** P ≤ 0.05, control vs. TAC; † P ≤ 0.05 difference between genotypes.

    Techniques Used: Mouse Assay, Activity Assay, Western Blot, Derivative Assay

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    Article Snippet: The supernatant was transferred to a new tube, and protein levels were quantitated using the BCA method (Thermo Fischer). .. Using SDS-PAGE (Nu-PAGE 4–12 % Bis-Tris protein gels) and Western blotting, 30–50 μg total protein were analyzed with the following antibodies: anti-phospho-AMP-activated protein kinase (AMPK) (Thr172) (Cell Signaling, #2531), anti-AMPK (Cell Signaling, #2532), anti-phospho-RAPTOR (Ser792) (Cell Signaling, #2083), anti-phospho-p38 (Thr180/Tyr182) (Cell Signaling, #4511), anti-p38 (Cell Signaling, #9212), anti-NRF2 (Novus biologicals, NB100-80011), anti-phospho-ERK1/2 (Thr202/Tyr204) (Cell Signaling, #4377), anti-ERK1/2 (Cell Signaling, #9102), anti-cleaved PARP (Asp214) (Cell Signaling, #9541), and anti-β-actin (Sigma), followed by secondary goat-anti-mouse (IRDye800CW) or secondary goat-anti-rabbit (IRDye680LT) antibodies. .. Antibody signals were detected and quantitated using a LI-COR instrument.

    Article Title: Receptors for Insulin-Like Growth Factor-2 and Androgens as Therapeutic Targets in Triple-Negative Breast Cancer
    Article Snippet: Using RIPA buffer, total protein was isolated, and protein concentration was determined using the Bradford assay. .. Forty micrograms of total cell protein were resolved by 4–15% SDS-PAGE, transferred to a PVDF membrane and probed with antibodies directed against IGF1R (1:500, Cell Signaling #3027), IR (1:500, Cell Signaling #3025), IGF2 (1:1000, AbCam ab9574), pAkt (1:1000, Cell Signaling #4060), MAPK (1:1000, Cell Signaling #9102), pMAPK (1:1000, Cell Signaling #4370), S6 (1:2000, Cell Signaling #2217), pS6 (1:2000, Cell Signaling #4858), and androgen receptor AR (1:500, Cell Signaling #5153). β-actin (1:2000, Sigma-Aldrich #A1978) was used as a loading control. .. Statistical Analysis GraphPad Prism software (Available online: www.graphpad.com ) was used for statistical analysis.

    Western Blot:

    Article Title: Targeting mitochondrial complex I using BAY 87-2243 reduces melanoma tumor growth
    Article Snippet: The supernatant was transferred to a new tube, and protein levels were quantitated using the BCA method (Thermo Fischer). .. Using SDS-PAGE (Nu-PAGE 4–12 % Bis-Tris protein gels) and Western blotting, 30–50 μg total protein were analyzed with the following antibodies: anti-phospho-AMP-activated protein kinase (AMPK) (Thr172) (Cell Signaling, #2531), anti-AMPK (Cell Signaling, #2532), anti-phospho-RAPTOR (Ser792) (Cell Signaling, #2083), anti-phospho-p38 (Thr180/Tyr182) (Cell Signaling, #4511), anti-p38 (Cell Signaling, #9212), anti-NRF2 (Novus biologicals, NB100-80011), anti-phospho-ERK1/2 (Thr202/Tyr204) (Cell Signaling, #4377), anti-ERK1/2 (Cell Signaling, #9102), anti-cleaved PARP (Asp214) (Cell Signaling, #9541), and anti-β-actin (Sigma), followed by secondary goat-anti-mouse (IRDye800CW) or secondary goat-anti-rabbit (IRDye680LT) antibodies. .. Antibody signals were detected and quantitated using a LI-COR instrument.

    other:

    Article Title: Activation of JNK Signaling Mediates Connective Tissue Growth Factor Expression and Scar Formation in Corneal Wound Healing
    Article Snippet: Antibodies against JNK, ERK1/2, p38 MAPK, phospho-JNK (Thr183/Tyr185), phospho-ERK1/2 (Thr202/Tyr204) and phospho-p38 MAPK (Thr180/Tyr182) were obtained from Cell Signaling Technology, Inc. (Danvers, MA).

    Article Title: Stimulation of JNK Phosphorylation by the PTTH in Prothoracic Glands of the Silkworm, Bombyx mori
    Article Snippet: Anti-phospho-SAPK/JNK (Thr183/Tyr185) (#9251), anti-phospho-ERK (#9101), anti-phospho-4E-BP (#9459), anti-total ERK (#9102), and anti-α-tubulin (#2144) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA).

    Lysis:

    Article Title: CD45RB Is a Novel Molecular Therapeutic Target to Inhibit A? Peptide-Induced Microglial MAPK Activation
    Article Snippet: Mouse IL-6 ELISA kit was obtained from eBioscience (San Diego, CA). .. Antibodies for phospho-p44/42 (pp44/42, Thr202/Tyr204) MAPK, phospho-p38 (pp38, Thr180/Tyr182) MAPK, total p44/42 MAPK, and total p38 MAPK were obtained from Cell Signaling Technology (Beverly, MA) as well as cell lysis buffer, and SDS blue loading buffers. .. PD98059 (a specific MEK1/2 inhibitor), SB203580 (a specific p38 MAPK inhibitor), and bisperoxo (1, 10-phenanthroline) oxovanadate (phen) were obtained from Calbiochem (La Jolla, CA).

    Incubation:

    Article Title: Extracellular Signal-Regulated Kinase 5 is Required for Low-Concentration H2O2-Induced Angiogenesis of Human Umbilical Vein Endothelial Cells
    Article Snippet: The protein concentration was measured using a BCA kit, and 30 μ g of protein per sample was used for polyacrylamide gel electrophoresis and then transferred to nitrocellulose membranes. .. The membranes were blocked with fat-free milk or 5% BSA for 1 h at room temperature and then incubated with various antibodies: ERK5 (#3372, Cell Signaling Technology, 1 : 1000), phosphorylated-ERK5 (#07-507, Millipore, 1 : 1000), MEF2C (#5030T, Cell Signaling Technology, 1 : 1000), phosphorylated-MEF2C (sc-377535, Santa Cruz, 1 : 200), ERK1/2 (#9102, Cell Signaling Technology, 1 : 1000), p-ERK1/2 (#9101, Cell Signaling Technology, 1 : 1000), and β -actin (ab8226, Abcam, 1 : 1000) overnight at 4°C. .. After the membranes were washed with TBST, they were incubated with HRP-linked secondary antibodies: goat anti-rabbit IgG (#7074, Cell Signaling Technology, 1 : 2000) or goat anti-mouse IgG (ab6728, Abcam, 1 : 2000) for 1 h at room temperature.

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  • 99
    Cell Signaling Technology Inc anti erk
    Pazopanib induces autophagic flux and ERK1/2 phosphorylation in BC cells. ( A ) Lysates from 5637 and J82 cells treated with 20 μ M pazopanib and 50 n M bafilomycin A1, alone or in combination, for 48 h were separated on 14% or 10% SDS–PAGE and probed with <t>anti-LC3</t> or anti-p62 and anti-GAPDH Abs, respectively. ( B ) Lysates from 5637 and J82 cells untreated or treated with 20 μ M pazopanib at different times were separated on 12% SDS–PAGE and probed with anti-pERK or <t>anti-ERK</t> Abs. Densitometric analysis, expressed as the mean±s.d., was performed by comparing treated with untreated cells, * P
    Anti Erk, 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/anti erk/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti erk - by Bioz Stars, 2021-06
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    97
    Cell Signaling Technology Inc anti total erk
    WDR 20 suppresses the phosphorylation of protein kinase B ( <t>AKT</t> ) and <t>ERK</t> in renal cell carcinoma cells. At 72 h after plating, the vector, 786O_ WDR 20 cl1, and 786O_ WDR 20 cl2 (A) or the vector, 786O_ WDR 20 cl2, and 786O_ WDR 20cl2wo (B) were subjected to Western blot analysis using antibodies against the phosphorylated form of AKT (Ser473), total AKT , the phosphorylated form of ERK (Thr202/Tyr204), total ERK , and GAPDH .
    Anti Total Erk, 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 erk/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti total erk - by Bioz Stars, 2021-06
    97/100 stars
      Buy from Supplier

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    Pazopanib induces autophagic flux and ERK1/2 phosphorylation in BC cells. ( A ) Lysates from 5637 and J82 cells treated with 20 μ M pazopanib and 50 n M bafilomycin A1, alone or in combination, for 48 h were separated on 14% or 10% SDS–PAGE and probed with anti-LC3 or anti-p62 and anti-GAPDH Abs, respectively. ( B ) Lysates from 5637 and J82 cells untreated or treated with 20 μ M pazopanib at different times were separated on 12% SDS–PAGE and probed with anti-pERK or anti-ERK Abs. Densitometric analysis, expressed as the mean±s.d., was performed by comparing treated with untreated cells, * P

    Journal: British Journal of Cancer

    Article Title: Pazopanib and sunitinib trigger autophagic and non-autophagic death of bladder tumour cells

    doi: 10.1038/bjc.2013.420

    Figure Lengend Snippet: Pazopanib induces autophagic flux and ERK1/2 phosphorylation in BC cells. ( A ) Lysates from 5637 and J82 cells treated with 20 μ M pazopanib and 50 n M bafilomycin A1, alone or in combination, for 48 h were separated on 14% or 10% SDS–PAGE and probed with anti-LC3 or anti-p62 and anti-GAPDH Abs, respectively. ( B ) Lysates from 5637 and J82 cells untreated or treated with 20 μ M pazopanib at different times were separated on 12% SDS–PAGE and probed with anti-pERK or anti-ERK Abs. Densitometric analysis, expressed as the mean±s.d., was performed by comparing treated with untreated cells, * P

    Article Snippet: The following rabbit polyclonal antibodies (Abs) were used: anti-p62 (1 : 1000, Cell Signaling Technology, Denver, CO, USA), anti-microtubule-associated protein-1 light chain 3 (LC3, 2 μ g ml−1 , Novus Biologicals, Littleton, CO, USA), anti-ERK (1 : 1000, Cell Signaling), and anti-cathepsin B (anti-CB, 1 : 1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

    Techniques: SDS Page

    MAP kinase components are required for efficient vacuolization, cell lysis, and virus release. ( a ) Inhibitors SP600125, Selumetinib, and SB203580 inhibit SV40-induced phosphorylation of JNK, ERK, and p38, respectively. CV-1 cells were infected at an MOI of 10. Inhibitor treatment was started at 12 h.p.i., and immunoblotting was performed on extracts prepared 48 h.p.i; ( b ) Bright-field images of CV-1 cells 48 h.p.i. after infection with SV40: CV-1 cells were infected at an MOI of 10 and treated with inhibitors against JNK, ERK, and p38 or DMSO vehicle at 12 h.p.i.. Scale bar, 20 µm; ( c ) The number of vacuolated cells two days after infection was quantified and normalized to DMSO-treated control cells. *** p

    Journal: Viruses

    Article Title: SV40 Polyomavirus Activates the Ras-MAPK Signaling Pathway for Vacuolization, Cell Death, and Virus Release

    doi: 10.3390/v12101128

    Figure Lengend Snippet: MAP kinase components are required for efficient vacuolization, cell lysis, and virus release. ( a ) Inhibitors SP600125, Selumetinib, and SB203580 inhibit SV40-induced phosphorylation of JNK, ERK, and p38, respectively. CV-1 cells were infected at an MOI of 10. Inhibitor treatment was started at 12 h.p.i., and immunoblotting was performed on extracts prepared 48 h.p.i; ( b ) Bright-field images of CV-1 cells 48 h.p.i. after infection with SV40: CV-1 cells were infected at an MOI of 10 and treated with inhibitors against JNK, ERK, and p38 or DMSO vehicle at 12 h.p.i.. Scale bar, 20 µm; ( c ) The number of vacuolated cells two days after infection was quantified and normalized to DMSO-treated control cells. *** p

    Article Snippet: The following primary antibodies were used for immunoblotting: anti-ß-actin (Abcam, #ab8227), anti-p-ERK (Cell Signaling Technology, CST, #4370), anti-p-JNK (CST, #4668), anti-p-p38 (CST, #4511), anti-p-MKK4 (CST, #9156), anti-ERK (CST, #4695), anti-JNK (CST, #9252), anti-p38 (CST, #8690), anti-p-ATF2 (CST, #9225), anti-p-cJun (CST, #3270), anti-large T antigen (PAb, #108), and anti-VP1 (PAb, #597).

    Techniques: Lysis, Infection

    SV40 infection activates intracellular signaling pathways at late times after infection. ( a ) Western blot analysis of phosphorylated and total p38, JNK, and ERK in mock-infected or SV40-infected CV-1 cells 48 h.p.i.: VP1 and beta-actin expression are shown as controls; ( b ) Western blot analysis of CV-1 cells over the time course of SV40 infection: Samples harvested at the indicated h.p.i. were analyzed for phosphorylated p38, JNK, ERK, MKK4, ATF2, and c-Jun as well as for beta-actin and VP1 expression.

    Journal: Viruses

    Article Title: SV40 Polyomavirus Activates the Ras-MAPK Signaling Pathway for Vacuolization, Cell Death, and Virus Release

    doi: 10.3390/v12101128

    Figure Lengend Snippet: SV40 infection activates intracellular signaling pathways at late times after infection. ( a ) Western blot analysis of phosphorylated and total p38, JNK, and ERK in mock-infected or SV40-infected CV-1 cells 48 h.p.i.: VP1 and beta-actin expression are shown as controls; ( b ) Western blot analysis of CV-1 cells over the time course of SV40 infection: Samples harvested at the indicated h.p.i. were analyzed for phosphorylated p38, JNK, ERK, MKK4, ATF2, and c-Jun as well as for beta-actin and VP1 expression.

    Article Snippet: The following primary antibodies were used for immunoblotting: anti-ß-actin (Abcam, #ab8227), anti-p-ERK (Cell Signaling Technology, CST, #4370), anti-p-JNK (CST, #4668), anti-p-p38 (CST, #4511), anti-p-MKK4 (CST, #9156), anti-ERK (CST, #4695), anti-JNK (CST, #9252), anti-p38 (CST, #8690), anti-p-ATF2 (CST, #9225), anti-p-cJun (CST, #3270), anti-large T antigen (PAb, #108), and anti-VP1 (PAb, #597).

    Techniques: Infection, Western Blot, Expressing

    Possible pathways for ABCA7-related LOAD pathogenesis. ABCA7 dysfunctions likely facilitate Aβ production in neurons by activating the SREBP2-BACE1 pathway directly and/or by modifying the brain lipid profile indirectly. Furthermore, ER stress is also induced by ABCA7 loss of function in neurons, which activates the PERK-eIF2α pathway, causing cognitive declines and further activation of the SREBP2-BACE1 pathway. Excessive activation of the ERK signaling pathway caused by ABCA7 dysfunctions may also contribute directly to AD pathogenesis.

    Journal: The Journal of Neuroscience

    Article Title: ABCA7 Deficiency Accelerates Amyloid-β Generation and Alzheimer's Neuronal Pathology

    doi: 10.1523/JNEUROSCI.3757-15.2016

    Figure Lengend Snippet: Possible pathways for ABCA7-related LOAD pathogenesis. ABCA7 dysfunctions likely facilitate Aβ production in neurons by activating the SREBP2-BACE1 pathway directly and/or by modifying the brain lipid profile indirectly. Furthermore, ER stress is also induced by ABCA7 loss of function in neurons, which activates the PERK-eIF2α pathway, causing cognitive declines and further activation of the SREBP2-BACE1 pathway. Excessive activation of the ERK signaling pathway caused by ABCA7 dysfunctions may also contribute directly to AD pathogenesis.

    Article Snippet: The following primary antibodies were used; anti-β-site APP cleaving enzyme 1 (anti-BACE1; Thermo Scientific), anti-APP C-terminal antibody (IBL), anti-82E1 antibody (IBL), anti-SREBP2 antibody (Cayman Chemical), anti-protein kinase R-like endoplasmic reticulum kinase (anti-PERK; Cell Signaling Technology), rabbit anti-phospho ERK1/2 (T202/Y204; Cell Signaling Technology, 4370), anti-ERK (Cell Signaling Technology), anti-phosphorylated eukaryotic initiation factor 2 (anti-eIF2; Cell Signaling Technology), anti-eIF2 (Cell Signaling Technology), anti-phospho-JNK (Cell Signaling Technology), anti-phospho-p38 (Cell Signaling Technology), anti-PS1 (Cell Signaling Technology), anti-GFAP antibody (Sigma-Aldrich), anti Iba1 antibody (WAKO), and anti-GAPDH (Sigma-Aldrich).

    Techniques: Activation Assay

    WDR 20 suppresses the phosphorylation of protein kinase B ( AKT ) and ERK in renal cell carcinoma cells. At 72 h after plating, the vector, 786O_ WDR 20 cl1, and 786O_ WDR 20 cl2 (A) or the vector, 786O_ WDR 20 cl2, and 786O_ WDR 20cl2wo (B) were subjected to Western blot analysis using antibodies against the phosphorylated form of AKT (Ser473), total AKT , the phosphorylated form of ERK (Thr202/Tyr204), total ERK , and GAPDH .

    Journal: Cancer Science

    Article Title: Downregulation of WDR20 due to loss of 14q is involved in the malignant transformation of clear cell renal cell carcinoma

    doi: 10.1111/cas.12892

    Figure Lengend Snippet: WDR 20 suppresses the phosphorylation of protein kinase B ( AKT ) and ERK in renal cell carcinoma cells. At 72 h after plating, the vector, 786O_ WDR 20 cl1, and 786O_ WDR 20 cl2 (A) or the vector, 786O_ WDR 20 cl2, and 786O_ WDR 20cl2wo (B) were subjected to Western blot analysis using antibodies against the phosphorylated form of AKT (Ser473), total AKT , the phosphorylated form of ERK (Thr202/Tyr204), total ERK , and GAPDH .

    Article Snippet: The primary antibodies were: anti‐FLAG M2 (Sigma‐Aldrich, St. Louis, MO, USA), anti‐p‐AKT S473 D9E, anti‐pan AKT, anti‐p‐ERK, anti‐total ERK (Cell Signaling Technology, Danvers, MA, USA), anti‐WDR20 (Bethyl, Montgomery, TX, USA), and anti‐GAPDH (Sigma‐Aldrich).

    Techniques: Plasmid Preparation, Western Blot