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  • 99
    Millipore monoclonal anti map kinase activated diphosphorylated erk 1 2 antibody
    Monoclonal Anti Map Kinase Activated Diphosphorylated Erk 1 2 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 714 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc erk1 2
    Interactions of ERK2 with mGluR1a in rat cerebellar neurons. a , b Immunoblot detection of mGluR1a ( a ) and mGluR5 ( b ) proteins in synaptic membranes extracted from the cerebellum and striatum. Note that a smaller amount of cerebellar proteins (2.5 µg) than that of striatal proteins (25 µg) was loaded ( a ). Major dimer bands around 250 kDa and weak monomer bands (130–140 kDa) were seen for mGluR1a in the cerebellum and striatum, while mGluR5 bands (mainly dimers) were seen in the striatum but not cerebellum. c, d Immunoblot detection of <t>ERK1/2</t> ( c ) and pERK1/2 ( d ) proteins in whole-cell homogenates and synaptic plasma membranes of cerebellar neurons (12.5 µg per lane). Note that ERK2 and pERK2 were expressed at a higher level than respective ERK1 and pERK1 in synaptic locations. e Coimmunoprecipitation ( IP ) assays of ERK1/2 and mGluR1a with the anti-mGluR1a antibody ( Ab ). Note that ERK2 clearly existed in mGluR1a precipitates ( lane 5 ). Lanes 3 and 4 showed no specific bands due to the lack of a precipitating antibody (L3) and the use of an irrelevant IgG (L4). f Reverse coimmunoprecipitation assays of ERK1/2 and mGluR1a with the anti-ERK1/2 antibody. g Coimmunoprecipitation of pERK1/2 and mGluR1a. Synaptic proteins solubilized from enriched synaptic plasma membranes of the rat cerebellum were used in above IP assays. Precipitated proteins were visualized by immunoblots ( IB ) with indicated antibodies
    Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 24006 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc p erk1 2
    The effects of AngII-Agtr1 inhibition on SMCs in aneurysmal lesions. ( A ) Changes in <t>p-ERK1/2</t> concentration in aneurysmal lesions after pharmacological intervention for the indicated time periods. Bars are 20 μm. ( B ) Western blot showing the effect
    P Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 8388 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti erk1 2
    GDF15 promotes oxidative metabolism and M2-like polarization. a Real-time PCR analysis of TGF-β RI ( Alk1–7 ) expression in BMDMs. b Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2 and 3. c Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2, SMAD3, <t>ERK1/2,</t> p38, and JNK, which was inhibited by the chemical inhibitors, SB431542 (an ALK4, 5, 7 inhibitor; 5 μM), PD98059 (an ERK1/2 inhibitor; 25 μM), SB203580 (a p38 inhibitor; 10 μM), and SP600125 (a JNK inhibitor; 10 μM). d , e OCR in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without SB431542 (5 μM). f , g Fatty acid oxidation in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without palmitate-BSA (200 μM). h Real-time PCR analysis of FAO-related genes in rGDF15 (300 ng/ml)-treated BMDMs. i – k Real-time PCR analysis of M1-related gene expression in rGDF15 (300 ng/ml)-treated BMDMs in the presence of IFN-γ (10 ng/ml) and LPS (100 ng/ml). ( l – n ) Real-time PCR analysis of M2-related gene expression in rGDF15-treated BMDMs in the presence of IL-4 (20 ng/ml). Data are expressed as means ± SEM and are representative of three independent experiments. * p
    Anti Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 8122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho erk1 2
    GDF15 promotes oxidative metabolism and M2-like polarization. a Real-time PCR analysis of TGF-β RI ( Alk1–7 ) expression in BMDMs. b Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2 and 3. c Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2, SMAD3, <t>ERK1/2,</t> p38, and JNK, which was inhibited by the chemical inhibitors, SB431542 (an ALK4, 5, 7 inhibitor; 5 μM), PD98059 (an ERK1/2 inhibitor; 25 μM), SB203580 (a p38 inhibitor; 10 μM), and SP600125 (a JNK inhibitor; 10 μM). d , e OCR in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without SB431542 (5 μM). f , g Fatty acid oxidation in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without palmitate-BSA (200 μM). h Real-time PCR analysis of FAO-related genes in rGDF15 (300 ng/ml)-treated BMDMs. i – k Real-time PCR analysis of M1-related gene expression in rGDF15 (300 ng/ml)-treated BMDMs in the presence of IFN-γ (10 ng/ml) and LPS (100 ng/ml). ( l – n ) Real-time PCR analysis of M2-related gene expression in rGDF15-treated BMDMs in the presence of IL-4 (20 ng/ml). Data are expressed as means ± SEM and are representative of three independent experiments. * p
    Phospho Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 9805 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology erk1 2
    Real-time PCR demonstrates that AGE-induced de novo α-SMA and loss of E-cadherin mRNA expression are regulated by RAGE through the <t>ERK1/2</t> MAP kinase pathway. Real-time PCR demonstrates that AGE-induced α-SMA ( a ) and reduced E-cadherin ( b ) mRNA expression by NRK52E cells at 24 hours are completely blocked by a neutralizing RAGE antibody (RAGE Ab) and a specific ERK1/2 MAP kinase inhibitor, PD 98059. CTL Ab, control antibody (rabbit IgG). Each bar represents the mean ± SD for three independent experiments. **, P
    Erk1 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 4426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phosphorylated erk1 2
    Real-time PCR demonstrates that AGE-induced de novo α-SMA and loss of E-cadherin mRNA expression are regulated by RAGE through the <t>ERK1/2</t> MAP kinase pathway. Real-time PCR demonstrates that AGE-induced α-SMA ( a ) and reduced E-cadherin ( b ) mRNA expression by NRK52E cells at 24 hours are completely blocked by a neutralizing RAGE antibody (RAGE Ab) and a specific ERK1/2 MAP kinase inhibitor, PD 98059. CTL Ab, control antibody (rabbit IgG). Each bar represents the mean ± SD for three independent experiments. **, P
    Phosphorylated Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 2640 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology erk1
    <t>ERK1</t> − / − ; ERK2 flox/flox ; Col2a1-Cre embryos. (A) Immunofluorescence using anti-ERK1 and ERK2 antibody showed reduced immunoreactivity in chondrocytes in the tibiae of ERK1 − / − ; ERK2 flox/flox ; Col2a1-Cre embryos at E18.5.
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    Santa Cruz Biotechnology anti erk1 2
    Expressions and prognostic roles of <t>MKP-4/ERK1/2</t> in HCC patients. a Western blot analysis revealed a lower expression of MKP-4 or higher expressions of ERK1/2, p-ERK1/2 in hepatocellular carcinoma (T) and adjacent non-tumorous tissues (N). b The bar chart demonstrates the ratio of MKP-4, ERK1/2 and p-ERK1/2 to GAPDH by quantitative analysis. * P
    Anti Erk1 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 1331 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam erk1 2
    Expressions and prognostic roles of <t>MKP-4/ERK1/2</t> in HCC patients. a Western blot analysis revealed a lower expression of MKP-4 or higher expressions of ERK1/2, p-ERK1/2 in hepatocellular carcinoma (T) and adjacent non-tumorous tissues (N). b The bar chart demonstrates the ratio of MKP-4, ERK1/2 and p-ERK1/2 to GAPDH by quantitative analysis. * P
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    Cell Signaling Technology Inc phospho p44 42 mapk
    Expressions and prognostic roles of <t>MKP-4/ERK1/2</t> in HCC patients. a Western blot analysis revealed a lower expression of MKP-4 or higher expressions of ERK1/2, p-ERK1/2 in hepatocellular carcinoma (T) and adjacent non-tumorous tissues (N). b The bar chart demonstrates the ratio of MKP-4, ERK1/2 and p-ERK1/2 to GAPDH by quantitative analysis. * P
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    Cell Signaling Technology Inc phospho erk1 2 thr202 tyr204
    Expressions and prognostic roles of <t>MKP-4/ERK1/2</t> in HCC patients. a Western blot analysis revealed a lower expression of MKP-4 or higher expressions of ERK1/2, p-ERK1/2 in hepatocellular carcinoma (T) and adjacent non-tumorous tissues (N). b The bar chart demonstrates the ratio of MKP-4, ERK1/2 and p-ERK1/2 to GAPDH by quantitative analysis. * P
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    Santa Cruz Biotechnology p erk1 2
    Effects of ApoSOD1 on Ca 2+ <t>/Akt/ERK1/2</t> signaling pathway in NSC-34 motor neurons. ( a ) (Top) Western blotting of SOD1 expression after inactivation with H 2 O 2 (200 mM) for several time points. (Bottom) Representative curve of the time-dependent inactivation of SOD1 in H 2 O 2 . ( b ) Bar graph depicting DCF-DA-detected ROS production in NSC-34 motor neurons exposed to chemical hypoxia in the absence and in the presence of SOD1 (400 ng/ml) or ApoSOD1 (400 ng/ml). Data are expressed as mean±S.E. of three different experiments. * P
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    Cell Signaling Technology Inc p44 42 mapk
    Effects of ApoSOD1 on Ca 2+ <t>/Akt/ERK1/2</t> signaling pathway in NSC-34 motor neurons. ( a ) (Top) Western blotting of SOD1 expression after inactivation with H 2 O 2 (200 mM) for several time points. (Bottom) Representative curve of the time-dependent inactivation of SOD1 in H 2 O 2 . ( b ) Bar graph depicting DCF-DA-detected ROS production in NSC-34 motor neurons exposed to chemical hypoxia in the absence and in the presence of SOD1 (400 ng/ml) or ApoSOD1 (400 ng/ml). Data are expressed as mean±S.E. of three different experiments. * P
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    Image Search Results


    Interactions of ERK2 with mGluR1a in rat cerebellar neurons. a , b Immunoblot detection of mGluR1a ( a ) and mGluR5 ( b ) proteins in synaptic membranes extracted from the cerebellum and striatum. Note that a smaller amount of cerebellar proteins (2.5 µg) than that of striatal proteins (25 µg) was loaded ( a ). Major dimer bands around 250 kDa and weak monomer bands (130–140 kDa) were seen for mGluR1a in the cerebellum and striatum, while mGluR5 bands (mainly dimers) were seen in the striatum but not cerebellum. c, d Immunoblot detection of ERK1/2 ( c ) and pERK1/2 ( d ) proteins in whole-cell homogenates and synaptic plasma membranes of cerebellar neurons (12.5 µg per lane). Note that ERK2 and pERK2 were expressed at a higher level than respective ERK1 and pERK1 in synaptic locations. e Coimmunoprecipitation ( IP ) assays of ERK1/2 and mGluR1a with the anti-mGluR1a antibody ( Ab ). Note that ERK2 clearly existed in mGluR1a precipitates ( lane 5 ). Lanes 3 and 4 showed no specific bands due to the lack of a precipitating antibody (L3) and the use of an irrelevant IgG (L4). f Reverse coimmunoprecipitation assays of ERK1/2 and mGluR1a with the anti-ERK1/2 antibody. g Coimmunoprecipitation of pERK1/2 and mGluR1a. Synaptic proteins solubilized from enriched synaptic plasma membranes of the rat cerebellum were used in above IP assays. Precipitated proteins were visualized by immunoblots ( IB ) with indicated antibodies

    Journal: Molecular neurobiology

    Article Title: Synaptic ERK2 Phosphorylates and Regulates Metabotropic Glutamate Receptor 1 In Vitro and in Neurons

    doi: 10.1007/s12035-016-0225-4

    Figure Lengend Snippet: Interactions of ERK2 with mGluR1a in rat cerebellar neurons. a , b Immunoblot detection of mGluR1a ( a ) and mGluR5 ( b ) proteins in synaptic membranes extracted from the cerebellum and striatum. Note that a smaller amount of cerebellar proteins (2.5 µg) than that of striatal proteins (25 µg) was loaded ( a ). Major dimer bands around 250 kDa and weak monomer bands (130–140 kDa) were seen for mGluR1a in the cerebellum and striatum, while mGluR5 bands (mainly dimers) were seen in the striatum but not cerebellum. c, d Immunoblot detection of ERK1/2 ( c ) and pERK1/2 ( d ) proteins in whole-cell homogenates and synaptic plasma membranes of cerebellar neurons (12.5 µg per lane). Note that ERK2 and pERK2 were expressed at a higher level than respective ERK1 and pERK1 in synaptic locations. e Coimmunoprecipitation ( IP ) assays of ERK1/2 and mGluR1a with the anti-mGluR1a antibody ( Ab ). Note that ERK2 clearly existed in mGluR1a precipitates ( lane 5 ). Lanes 3 and 4 showed no specific bands due to the lack of a precipitating antibody (L3) and the use of an irrelevant IgG (L4). f Reverse coimmunoprecipitation assays of ERK1/2 and mGluR1a with the anti-ERK1/2 antibody. g Coimmunoprecipitation of pERK1/2 and mGluR1a. Synaptic proteins solubilized from enriched synaptic plasma membranes of the rat cerebellum were used in above IP assays. Precipitated proteins were visualized by immunoblots ( IB ) with indicated antibodies

    Article Snippet: Antibodies used in this study include rabbit antibodies against mGluR1a (Millipore; 1:2000), ERK1/2 (Cell Signaling; 1:2000), pERK1/2 (phosphorylation at T185/Y187 for pERK2; Cell Signaling; 1:1000), GST (Sigma; 1:2000), a phospho-serine/ threonine-proline motif (pS/TP, Abcam, Cambridge, MA; 1:1000), a phospho-MAPK motif PXpSP or pSPXR/K (Cell Signaling; 1:1000), phosphoserine (Invitrogen; 1:1000), phosphothreonine (Invitrogen; 1:1000), Src with phosphorylated tyrosine 416 (pY416, Cell Signaling; 1:1000), Src (Cell Signaling; 1:2000), or actin (Sigma; 1:4000) or mouse antibodies against mGluR1a (BD, Franklin Lakes, NJ; 1:2000), ERK1/2 (Cell Signaling; 1:2000), ERK2 (Abcam; 1:2000), or a phosphothreonine-proline motif (pTP, Cell Signaling; 1:1000).

    Techniques: Western Blot

    ERK-mediated phosphorylation of mGluR1a in rat cerebellar neurons. a–c Phosphorylation of mGluR1a at ERK-preferred motifs, including S/TP ( a ), TP ( b ), and PXSP ( c ). Cerebellar mGluR1a was immunopurified by the anti-mGluR1a antibody. Phosphorylation signals at three phosphomotifs (pS/TP, pTP, and PXpSP) in immunopurified mGluR1a were detected by immunoblots ( IB ) with indicated antibodies. d Dephosphorylation of mGluR1a PXSP phosphorylation by λ-protein phosphatase (λ-PP). Cerebellar slices were incubated with λ-PP (200–400 units) for 1 h at 30 °C. Note that PXSP phosphorylation was dephosphorylated by λ-PP. e Representative immunoblots illustrating effects of U0126 on phosphorylation of mGluR1a at PXSP and TP motifs and on pERK1/2-mGluR1a and ERK1/2-mGluR1a interactions in cerebellar neurons. f, g Quantifications of effects of U0126 on mGluR1a PXSP ( f ) and TP ( g ) phosphorylation. h Quantifications of effects of U0126 on pERK2-mGluR1a and ERK2-mGluR1a interactions. Cerebellar slices were incubated with U0126 (0.5 or 5 µM) for 30 min at 30 °C ( e–h ). Slices were collected for mGluR1a immunoprecipitation ( IP ). Phosphorylation levels of mGluR1a at PXSP and TP and proteins bound to immunopurified mGluR1a (pERK1/2 and ERK1/2) were visualized by immunoblots. Values in graphs were measured as ratios of PXpSP to mGluR1a ( f ), pTP to mGluR1a ( g ), pERK2 to mGluR1a ( h ), and ERK2 to mGluR1a ( h ). All values were analyzed by one-way ANOVA: PXpSP ( f ), F (2, 7) = 5.05, p

    Journal: Molecular neurobiology

    Article Title: Synaptic ERK2 Phosphorylates and Regulates Metabotropic Glutamate Receptor 1 In Vitro and in Neurons

    doi: 10.1007/s12035-016-0225-4

    Figure Lengend Snippet: ERK-mediated phosphorylation of mGluR1a in rat cerebellar neurons. a–c Phosphorylation of mGluR1a at ERK-preferred motifs, including S/TP ( a ), TP ( b ), and PXSP ( c ). Cerebellar mGluR1a was immunopurified by the anti-mGluR1a antibody. Phosphorylation signals at three phosphomotifs (pS/TP, pTP, and PXpSP) in immunopurified mGluR1a were detected by immunoblots ( IB ) with indicated antibodies. d Dephosphorylation of mGluR1a PXSP phosphorylation by λ-protein phosphatase (λ-PP). Cerebellar slices were incubated with λ-PP (200–400 units) for 1 h at 30 °C. Note that PXSP phosphorylation was dephosphorylated by λ-PP. e Representative immunoblots illustrating effects of U0126 on phosphorylation of mGluR1a at PXSP and TP motifs and on pERK1/2-mGluR1a and ERK1/2-mGluR1a interactions in cerebellar neurons. f, g Quantifications of effects of U0126 on mGluR1a PXSP ( f ) and TP ( g ) phosphorylation. h Quantifications of effects of U0126 on pERK2-mGluR1a and ERK2-mGluR1a interactions. Cerebellar slices were incubated with U0126 (0.5 or 5 µM) for 30 min at 30 °C ( e–h ). Slices were collected for mGluR1a immunoprecipitation ( IP ). Phosphorylation levels of mGluR1a at PXSP and TP and proteins bound to immunopurified mGluR1a (pERK1/2 and ERK1/2) were visualized by immunoblots. Values in graphs were measured as ratios of PXpSP to mGluR1a ( f ), pTP to mGluR1a ( g ), pERK2 to mGluR1a ( h ), and ERK2 to mGluR1a ( h ). All values were analyzed by one-way ANOVA: PXpSP ( f ), F (2, 7) = 5.05, p

    Article Snippet: Antibodies used in this study include rabbit antibodies against mGluR1a (Millipore; 1:2000), ERK1/2 (Cell Signaling; 1:2000), pERK1/2 (phosphorylation at T185/Y187 for pERK2; Cell Signaling; 1:1000), GST (Sigma; 1:2000), a phospho-serine/ threonine-proline motif (pS/TP, Abcam, Cambridge, MA; 1:1000), a phospho-MAPK motif PXpSP or pSPXR/K (Cell Signaling; 1:1000), phosphoserine (Invitrogen; 1:1000), phosphothreonine (Invitrogen; 1:1000), Src with phosphorylated tyrosine 416 (pY416, Cell Signaling; 1:1000), Src (Cell Signaling; 1:2000), or actin (Sigma; 1:4000) or mouse antibodies against mGluR1a (BD, Franklin Lakes, NJ; 1:2000), ERK1/2 (Cell Signaling; 1:2000), ERK2 (Abcam; 1:2000), or a phosphothreonine-proline motif (pTP, Cell Signaling; 1:1000).

    Techniques: Western Blot, De-Phosphorylation Assay, Incubation, Immunoprecipitation

    ( A . (b) Densitometry analysis determined from western blots. Phosphorylated (p-ERK1/2) relative to total (t-ERK1/2). Values represent mean ± SD ( n = 3). * represents significant difference compared to 0 h by student’s t test, p

    Journal: International Journal of Molecular Sciences

    Article Title: Chlorogenic Acid (CGA) Isomers Alleviate Interleukin 8 (IL-8) Production in Caco-2 Cells by Decreasing Phosphorylation of p38 and Increasing Cell Integrity

    doi: 10.3390/ijms19123873

    Figure Lengend Snippet: ( A . (b) Densitometry analysis determined from western blots. Phosphorylated (p-ERK1/2) relative to total (t-ERK1/2). Values represent mean ± SD ( n = 3). * represents significant difference compared to 0 h by student’s t test, p

    Article Snippet: Specifically, the membrane that was used for the detection of p-Erk1/2 was washed three times with TBST and incubated in a mixture antibody solution that contained both anti-phospho-Erk1/2 (phospho Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (1:2000 dilution) (4370S, Cell Signaling Technology, Danvers, MA, USA) and anti-Erk1/2 (137F5) Rabbit mAb (1:1000 dilution) (4695S, Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Western Blot

    The GluN2B CaMKII Binding Site Is Dispensable for Theta Burst LTP (A–C) Activity-dependent signaling to ERK1/2 does not require GluN2B CTD-specific sequences. DIV9 cortical neurons of the indicated genotypes were treated with TTX (500 nM), KN-62 (10 μM), or MK-801 (10 μM) for 1 hr, after which protein extracts were made and subjected to western blot analysis for phospho-ERK1/2 levels, normalized to total ERK1/2. (A) shows quantitation and (B) and (C) show example blots. ∗ p

    Journal: Cell Reports

    Article Title: The Developmental Shift of NMDA Receptor Composition Proceeds Independently of GluN2 Subunit-Specific GluN2 C-Terminal Sequences

    doi: 10.1016/j.celrep.2018.09.089

    Figure Lengend Snippet: The GluN2B CaMKII Binding Site Is Dispensable for Theta Burst LTP (A–C) Activity-dependent signaling to ERK1/2 does not require GluN2B CTD-specific sequences. DIV9 cortical neurons of the indicated genotypes were treated with TTX (500 nM), KN-62 (10 μM), or MK-801 (10 μM) for 1 hr, after which protein extracts were made and subjected to western blot analysis for phospho-ERK1/2 levels, normalized to total ERK1/2. (A) shows quantitation and (B) and (C) show example blots. ∗ p

    Article Snippet: The membranes were incubated at 4°C overnight with the primary antibodies diluted in blocking solution: anti phospho-(Ser-1303) GluN2B (1: 2000, Millipore), anti-phospho (Tyr-1472) GluN2B (1:2000, Millipore), anti-phospho (Ser1480) GluN2B (1:2000, Abcam), anti-GluN2B (C terminus, 1:4000, BD Transduction Laboratories), anti-GluN2B (N terminus 1:2000, Thermo Fisher Scientific), anti-GluN2A (N terminus, 1:1000, Thermo Fisher Scientific), anti-CamKiiα (1:8000, Thermo Fisher Scientific), anti-ERK1/2 (1:2000, Cell Signaling), anti-phospho ERK1/2 (1:2000, Cell Signaling) and anti-beta actin (1:200000, Abcam).

    Techniques: Binding Assay, Activity Assay, Western Blot, Quantitation Assay

    LA inhibited P38 MAPK, calpain1/CDK5 and GSK3β pathways but promoted PP2A. (A) Western blotting analysis of p-ERK1/2, ERK1/2, p-JNK1/2, JNK1/2, p-P38, and P38. β-actin served as the loading control. ( B-D) Quantitative analysis of p-ERK1/2, p-JNK1/2 and p-P38. ( E) Western blot detection of calpain1, p-CDK5, CDK5 and P35/25 in the brains. ( F-H) Quantification of calpain1, p-CDK5 and the P25/P35 ratio. (I) Western blot for p-GSK3α/β, GSK3α/β and PP2A expression. ( J-L) Quantitative analysis of p-GSK3α, p-GSK3β and PP2A. All results are presented as the mean ± SEM (n = 7). *p

    Journal: Redox Biology

    Article Title: α-Lipoic acid improves abnormal behavior by mitigation of oxidative stress, inflammation, ferroptosis, and tauopathy in P301S Tau transgenic mice

    doi: 10.1016/j.redox.2017.11.001

    Figure Lengend Snippet: LA inhibited P38 MAPK, calpain1/CDK5 and GSK3β pathways but promoted PP2A. (A) Western blotting analysis of p-ERK1/2, ERK1/2, p-JNK1/2, JNK1/2, p-P38, and P38. β-actin served as the loading control. ( B-D) Quantitative analysis of p-ERK1/2, p-JNK1/2 and p-P38. ( E) Western blot detection of calpain1, p-CDK5, CDK5 and P35/25 in the brains. ( F-H) Quantification of calpain1, p-CDK5 and the P25/P35 ratio. (I) Western blot for p-GSK3α/β, GSK3α/β and PP2A expression. ( J-L) Quantitative analysis of p-GSK3α, p-GSK3β and PP2A. All results are presented as the mean ± SEM (n = 7). *p

    Article Snippet: The primary antibodies used in the study were as follows: mouse-anti-apoptosis inducing factor (AIF, 1:1000, Santa Cruz), mouse-anti-Bcl2 (1:1000, Santa Cruz), rabbit-anti-Bax (1:1000, Santa Cruz), mouse-anti-Beclin1 (1:1000, ImmunoWay), rabbit-anti-calpain1 (1:2000, Abcam), rabbit-anti-p-cyclin dependent kinase 5 (Tyr15) (CDK5, 1:2000, Sigma), rabbit-anti-CDK5 (1:2000, Abcam), rabbit-anti-caspase3 (1:2000, CST), rabbit-anti-divalent mental transporter 1 (DMT1, 1:1000, Alpha diagnostic), rabbit-anti-p-extracellular regulated protein kinases1/2 (Thr202/Thr204) (ERK1/2, 1:2000, CST), rabbit-anti-ERK1/2 (1:2000, CST), goat-anti-ferroportin 1 (Fpn 1, 1:1000, Santa Cruz), rabbit-anti-glial fibrillary acidic protein (GFAP, 1:1000, Sigma), rabbit-anti-glutathione peroxidase 4 (GPx4, 1:1000, Santa Cruz), rabbit-anti-p-glycogen synthase kinase 3 alpha/βeta (Ser21/9) (p-GSK3α/β, 1:2000, CST), rabbit-anti-GSK3α/β (1:2000, CST), rabbit-anti-ionized binding adaptor protein 1 (Iba1, 1:1000, Abcam), rabbit-anti-interleukin 1 beta (IL-1β, 1:1000, Santa Cruz), mouse-anti-p-SAPK/JNK (Thr183/Tyr185) (1:2000, CST), rabbit-anti-SAPK/JNK (1:2000, CST), rabbit-anti-LC3A/B (1:2000, CST), rabbit-anti-NeuN (1:2000, CST), rabbit-anti-postsynaptic density protein 95 (PSD95, 1:2000, CST), rabbit-anti-p35/25 (1:2000, CST), rabbit-anti-protein phosphatase 2A C subunit (PP2A, 1:2000, CST), rabbit-anti-p-P38 mitogen-activated protein kinase (Thr180/Tyr182) (p-P38 MAPK, 1:2000, CST), rabbit-anti-P38 MAPK (1:2000, CST), rabbit-anti-superoxide dismutase 1 (SOD1, 1:1000, CST), mouse-anti-synaptophysin (SYP, 1:2000, Sigma), mouse-anti-tumor necrosis factor alpha (TNFα, 1:500, Santa Cruz), mouse-anti-transferrin receptor (TFR, 1:1000, Invitrogen), mouse-anti-Tau (Tau46) (1:2000, CST), rabbit-anti-p-Tau (Thr181) (1:2000, CST), rabbit-anti-p-Tau (Ser416) (1:2000, CST), rabbit-anti-p-Tau (Ser202) (1:2000, CST), rabbit-anti-p-Tau (Ser396) (1:1000, Sigma), rabbit-anti-p-Tau (Ser404) (1:2000, Sigma), rabbit-anti-p-Tau (Thr231) (1:1000, Sigma), rabbit-anti-cystine-glutamate antiporter (xCT, 1:2000, Abcam), and mouse-anti-β-actin (1:10000, Sigma).

    Techniques: Western Blot, Expressing

    Inhibitory effect of pep234 on AjKissR1 and AjKissR2 activation. ( A ) Intracellular Ca 2+ mobilization in AjKissR1- and AjKissR2-expressing HEK293 cells was measured in response to 100 nM AjKiss1a or AjKiss1b-10 pre-treated with DMSO or the KISS1 antagonist pep234 (1.0 μM). ( B ) ERK1/2 phosphorylation activity of kisspeptins and the inhibitory effect of pep234 in AjKissR1- and AjKissR2-expressing HEK293 cells. Samples were measured after 2 hr of ligand administration with or without pep234 pre-treatment. Error bars represent SEM for three independent experiments. Immunoblots were quantified using a Bio-Rad Quantity One Imaging system. All data shown are representative of at least three independent experiments.

    Journal: eLife

    Article Title: Existence and functions of a kisspeptin neuropeptide signaling system in a non-chordate deuterostome species

    doi: 10.7554/eLife.53370

    Figure Lengend Snippet: Inhibitory effect of pep234 on AjKissR1 and AjKissR2 activation. ( A ) Intracellular Ca 2+ mobilization in AjKissR1- and AjKissR2-expressing HEK293 cells was measured in response to 100 nM AjKiss1a or AjKiss1b-10 pre-treated with DMSO or the KISS1 antagonist pep234 (1.0 μM). ( B ) ERK1/2 phosphorylation activity of kisspeptins and the inhibitory effect of pep234 in AjKissR1- and AjKissR2-expressing HEK293 cells. Samples were measured after 2 hr of ligand administration with or without pep234 pre-treatment. Error bars represent SEM for three independent experiments. Immunoblots were quantified using a Bio-Rad Quantity One Imaging system. All data shown are representative of at least three independent experiments.

    Article Snippet: Blots were stripped and reprobed using anti-ERK1/2 antibody (1:2000; Cell Signaling Technology) as a control for protein loading.

    Techniques: Activation Assay, Expressing, Activity Assay, Western Blot, Imaging

    The effects of AngII-Agtr1 inhibition on SMCs in aneurysmal lesions. ( A ) Changes in p-ERK1/2 concentration in aneurysmal lesions after pharmacological intervention for the indicated time periods. Bars are 20 μm. ( B ) Western blot showing the effect

    Journal: Science translational medicine

    Article Title: Angiotensin converting enzyme-induced activation of local angiotensin signaling is required for ascending aortic aneurysms in fibulin-4 deficient mice

    doi: 10.1126/scitranslmed.3005025

    Figure Lengend Snippet: The effects of AngII-Agtr1 inhibition on SMCs in aneurysmal lesions. ( A ) Changes in p-ERK1/2 concentration in aneurysmal lesions after pharmacological intervention for the indicated time periods. Bars are 20 μm. ( B ) Western blot showing the effect

    Article Snippet: Immunostaining was performed with antibodies for ACE (1:100, Santa Cruz) and p-ERK1/2 (1:100, Cell Signaling) as previously described ( ).

    Techniques: Inhibition, Concentration Assay, Western Blot

    GDF15 promotes oxidative metabolism and M2-like polarization. a Real-time PCR analysis of TGF-β RI ( Alk1–7 ) expression in BMDMs. b Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2 and 3. c Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2, SMAD3, ERK1/2, p38, and JNK, which was inhibited by the chemical inhibitors, SB431542 (an ALK4, 5, 7 inhibitor; 5 μM), PD98059 (an ERK1/2 inhibitor; 25 μM), SB203580 (a p38 inhibitor; 10 μM), and SP600125 (a JNK inhibitor; 10 μM). d , e OCR in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without SB431542 (5 μM). f , g Fatty acid oxidation in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without palmitate-BSA (200 μM). h Real-time PCR analysis of FAO-related genes in rGDF15 (300 ng/ml)-treated BMDMs. i – k Real-time PCR analysis of M1-related gene expression in rGDF15 (300 ng/ml)-treated BMDMs in the presence of IFN-γ (10 ng/ml) and LPS (100 ng/ml). ( l – n ) Real-time PCR analysis of M2-related gene expression in rGDF15-treated BMDMs in the presence of IL-4 (20 ng/ml). Data are expressed as means ± SEM and are representative of three independent experiments. * p

    Journal: Nature Communications

    Article Title: Reduced oxidative capacity in macrophages results in systemic insulin resistance

    doi: 10.1038/s41467-018-03998-z

    Figure Lengend Snippet: GDF15 promotes oxidative metabolism and M2-like polarization. a Real-time PCR analysis of TGF-β RI ( Alk1–7 ) expression in BMDMs. b Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2 and 3. c Immunoblot analysis of rGDF15-induced phosphorylation of SMAD2, SMAD3, ERK1/2, p38, and JNK, which was inhibited by the chemical inhibitors, SB431542 (an ALK4, 5, 7 inhibitor; 5 μM), PD98059 (an ERK1/2 inhibitor; 25 μM), SB203580 (a p38 inhibitor; 10 μM), and SP600125 (a JNK inhibitor; 10 μM). d , e OCR in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without SB431542 (5 μM). f , g Fatty acid oxidation in BMDMs treated with rGDF15 (300 ng/ml) for 24 h with or without palmitate-BSA (200 μM). h Real-time PCR analysis of FAO-related genes in rGDF15 (300 ng/ml)-treated BMDMs. i – k Real-time PCR analysis of M1-related gene expression in rGDF15 (300 ng/ml)-treated BMDMs in the presence of IFN-γ (10 ng/ml) and LPS (100 ng/ml). ( l – n ) Real-time PCR analysis of M2-related gene expression in rGDF15-treated BMDMs in the presence of IL-4 (20 ng/ml). Data are expressed as means ± SEM and are representative of three independent experiments. * p

    Article Snippet: Anti-phospho-STAT6, anti-STAT6, anti-phospho-SMAD2 (#3104), anti-SMAD2, anti-phospho-SMAD3, anti-SMAD3, anti-phospho-SMAD1, 5, and 9, anti-SMAD1, 5, and 9, anti-phospho-ERK1/2 (44/42 MAPK) (#4370), anti-ERK1/2 (#9102), anti-phospho-p38, anti-p38, anti-phospho-JNK, anti-JNK, and anti-SDHA (#5893) antibodies were purchased from Cell Signaling Technology (Beverley, MA, USA).

    Techniques: Real-time Polymerase Chain Reaction, Expressing

    Protein assay. Expression of phosphorylated (A) Akt, (B) STAT3, and (C) ERK1/2 in normal and nonalcoholic steatohepatitis liver groups 6 h after PH. Expression levels were detected by western blot analysis. Ratios of densitometry were calculated by each score/score of control 70% PH (D). Data are expressed as the mean ± SD. n = 5-7 per groups. b P

    Journal: World Journal of Gastroenterology

    Article Title: Evaluation of safety for hepatectomy in a novel mouse model with nonalcoholic-steatohepatitis

    doi: 10.3748/wjg.v24.i15.1622

    Figure Lengend Snippet: Protein assay. Expression of phosphorylated (A) Akt, (B) STAT3, and (C) ERK1/2 in normal and nonalcoholic steatohepatitis liver groups 6 h after PH. Expression levels were detected by western blot analysis. Ratios of densitometry were calculated by each score/score of control 70% PH (D). Data are expressed as the mean ± SD. n = 5-7 per groups. b P

    Article Snippet: Immunoblots were developed using polyclonal antibodies against phospho-AKT (9271), total AKT (9272), phospho-STAT3 (9131), total STAT3 (9132), phospho-ERK1/2 (9101), and total ERK1/2 (9102) (Cell Signaling Technology, Beverly, MA, United States).

    Techniques: Expressing, Western Blot

    Real-time PCR demonstrates that AGE-induced de novo α-SMA and loss of E-cadherin mRNA expression are regulated by RAGE through the ERK1/2 MAP kinase pathway. Real-time PCR demonstrates that AGE-induced α-SMA ( a ) and reduced E-cadherin ( b ) mRNA expression by NRK52E cells at 24 hours are completely blocked by a neutralizing RAGE antibody (RAGE Ab) and a specific ERK1/2 MAP kinase inhibitor, PD 98059. CTL Ab, control antibody (rabbit IgG). Each bar represents the mean ± SD for three independent experiments. **, P

    Journal: The American Journal of Pathology

    Article Title: Advanced Glycation End Products Induce Tubular Epithelial-Myofibroblast Transition through the RAGE-ERK1/2 MAP Kinase Signaling Pathway

    doi:

    Figure Lengend Snippet: Real-time PCR demonstrates that AGE-induced de novo α-SMA and loss of E-cadherin mRNA expression are regulated by RAGE through the ERK1/2 MAP kinase pathway. Real-time PCR demonstrates that AGE-induced α-SMA ( a ) and reduced E-cadherin ( b ) mRNA expression by NRK52E cells at 24 hours are completely blocked by a neutralizing RAGE antibody (RAGE Ab) and a specific ERK1/2 MAP kinase inhibitor, PD 98059. CTL Ab, control antibody (rabbit IgG). Each bar represents the mean ± SD for three independent experiments. **, P

    Article Snippet: Specifically, AGEs are able to activate ERK1/2 within 15 minutes and this is blocked by a soluble truncated receptor for AGE, sRAGE, and an inhibitor to ERK1/2, PD98059.

    Techniques: Real-time Polymerase Chain Reaction, Expressing, CTL Assay

    Western blot analysis demonstrates that AGE-induced loss of E-cadherin protein expression is TGF-β-independent and RAGE-ERK-dependent. Results show that AGE-induced loss of E-cadherin expression by NRK52E cells at 24 hours is not inhibited by a neutralizing TGF-β antibody (A), but is prevented by addition of sRAGE and an ERK1/2 inhibitor, PD98059. The specificity of a neutralizing anti-TGF-β antibody to block TGF-β-induced loss of E-cadherin is demonstrated in the last two lanes . CTL Ab, control rabbit antibody (IgG). Each bar represents the mean ± SD for four independent experiments. *, P

    Journal: The American Journal of Pathology

    Article Title: Advanced Glycation End Products Induce Tubular Epithelial-Myofibroblast Transition through the RAGE-ERK1/2 MAP Kinase Signaling Pathway

    doi:

    Figure Lengend Snippet: Western blot analysis demonstrates that AGE-induced loss of E-cadherin protein expression is TGF-β-independent and RAGE-ERK-dependent. Results show that AGE-induced loss of E-cadherin expression by NRK52E cells at 24 hours is not inhibited by a neutralizing TGF-β antibody (A), but is prevented by addition of sRAGE and an ERK1/2 inhibitor, PD98059. The specificity of a neutralizing anti-TGF-β antibody to block TGF-β-induced loss of E-cadherin is demonstrated in the last two lanes . CTL Ab, control rabbit antibody (IgG). Each bar represents the mean ± SD for four independent experiments. *, P

    Article Snippet: Specifically, AGEs are able to activate ERK1/2 within 15 minutes and this is blocked by a soluble truncated receptor for AGE, sRAGE, and an inhibitor to ERK1/2, PD98059.

    Techniques: Western Blot, Expressing, Blocking Assay, CTL Assay

    Double immunocytochemistry demonstrates that AGEs induce TEMT as determined by de novo expression of α-SMA and a partial loss of E-cadherin in NRK52E cells. NRK52E cells were stimulated with control BSA (30 μg/ml, A ) or AGE-BSA (30 μg/ml, B–F ) for 24 hours in the presence of an isotype control normal rabbit IgG (10 μg/ml, C ), a neutralizing TGF-β antibody (10 μg/ml, D ), a neutralizing RAGE antibody (10 μg/ml, E ), or a specific ERK1/2 MAP kinase inhibitor (PD 98059, 10 μmol/L, F ). Cells were stained with mAbs to α-SMA (myofibroblasts, red) and E-cadherin (epithelial cells, blue). The percentage of transformed cells (α-SMA+) is shown in ( G ). Results show that AGE-induced α-SMA expression in TECs is blocked by a neutralizing RAGE and an ERK1/2 MAP kinase inhibitor, but not by a neutralizing TGF-β antibody. Tab, anti-TGF-β antibody; Cab, control antibody; Rab, anti-RAGE antibody; PD, PD 98059. Data represent the mean ± SD for five experiments. Nuclei are stained with hematoxylin. Magnification, ×250. ***, P

    Journal: The American Journal of Pathology

    Article Title: Advanced Glycation End Products Induce Tubular Epithelial-Myofibroblast Transition through the RAGE-ERK1/2 MAP Kinase Signaling Pathway

    doi:

    Figure Lengend Snippet: Double immunocytochemistry demonstrates that AGEs induce TEMT as determined by de novo expression of α-SMA and a partial loss of E-cadherin in NRK52E cells. NRK52E cells were stimulated with control BSA (30 μg/ml, A ) or AGE-BSA (30 μg/ml, B–F ) for 24 hours in the presence of an isotype control normal rabbit IgG (10 μg/ml, C ), a neutralizing TGF-β antibody (10 μg/ml, D ), a neutralizing RAGE antibody (10 μg/ml, E ), or a specific ERK1/2 MAP kinase inhibitor (PD 98059, 10 μmol/L, F ). Cells were stained with mAbs to α-SMA (myofibroblasts, red) and E-cadherin (epithelial cells, blue). The percentage of transformed cells (α-SMA+) is shown in ( G ). Results show that AGE-induced α-SMA expression in TECs is blocked by a neutralizing RAGE and an ERK1/2 MAP kinase inhibitor, but not by a neutralizing TGF-β antibody. Tab, anti-TGF-β antibody; Cab, control antibody; Rab, anti-RAGE antibody; PD, PD 98059. Data represent the mean ± SD for five experiments. Nuclei are stained with hematoxylin. Magnification, ×250. ***, P

    Article Snippet: Specifically, AGEs are able to activate ERK1/2 within 15 minutes and this is blocked by a soluble truncated receptor for AGE, sRAGE, and an inhibitor to ERK1/2, PD98059.

    Techniques: Immunocytochemistry, Expressing, Staining, Transformation Assay

    Western blot analysis demonstrates that AGE-induced α-SMA protein expression is TGF-β-independent, but RAGE-ERK-dependent. Results show that AGE-induced α-SMA expression by NRK52E cells at 24 hours is not inhibited by a neutralizing TGF-β antibody, but is completely blocked by sRAGE and an ERK1/2 inhibitor, PD98059. The specificity of a neutralizing anti-TGF-β antibody to block TGF-β-induced α-SMA expression is shown in the last two lanes . CTL Ab, control rabbit antibody (IgG). Each bar represents the mean ± SD for three independent experiments. *, P

    Journal: The American Journal of Pathology

    Article Title: Advanced Glycation End Products Induce Tubular Epithelial-Myofibroblast Transition through the RAGE-ERK1/2 MAP Kinase Signaling Pathway

    doi:

    Figure Lengend Snippet: Western blot analysis demonstrates that AGE-induced α-SMA protein expression is TGF-β-independent, but RAGE-ERK-dependent. Results show that AGE-induced α-SMA expression by NRK52E cells at 24 hours is not inhibited by a neutralizing TGF-β antibody, but is completely blocked by sRAGE and an ERK1/2 inhibitor, PD98059. The specificity of a neutralizing anti-TGF-β antibody to block TGF-β-induced α-SMA expression is shown in the last two lanes . CTL Ab, control rabbit antibody (IgG). Each bar represents the mean ± SD for three independent experiments. *, P

    Article Snippet: Specifically, AGEs are able to activate ERK1/2 within 15 minutes and this is blocked by a soluble truncated receptor for AGE, sRAGE, and an inhibitor to ERK1/2, PD98059.

    Techniques: Western Blot, Expressing, Blocking Assay, CTL Assay

    Western blot analysis demonstrates that AGEs signal through RAGE to activate the ERK1/2 signaling pathway. a: A representative Western blot shows that AGE-BSA (30 μg/ml) induces ERK1/2 phosphorylation in NRK52E in a time-dependent manner, being significant at 15 minutes. b: AGEs, but not BSA, induce ERK1/2 phosphorylation (at 30 minutes) in a dose-dependent manner and this is blocked by both sRAGE and a specific ERK1/2 inhibitor, PD 98059. Each sample represents the results from three independent experiments.

    Journal: The American Journal of Pathology

    Article Title: Advanced Glycation End Products Induce Tubular Epithelial-Myofibroblast Transition through the RAGE-ERK1/2 MAP Kinase Signaling Pathway

    doi:

    Figure Lengend Snippet: Western blot analysis demonstrates that AGEs signal through RAGE to activate the ERK1/2 signaling pathway. a: A representative Western blot shows that AGE-BSA (30 μg/ml) induces ERK1/2 phosphorylation in NRK52E in a time-dependent manner, being significant at 15 minutes. b: AGEs, but not BSA, induce ERK1/2 phosphorylation (at 30 minutes) in a dose-dependent manner and this is blocked by both sRAGE and a specific ERK1/2 inhibitor, PD 98059. Each sample represents the results from three independent experiments.

    Article Snippet: Specifically, AGEs are able to activate ERK1/2 within 15 minutes and this is blocked by a soluble truncated receptor for AGE, sRAGE, and an inhibitor to ERK1/2, PD98059.

    Techniques: Western Blot

    Western blot analysis shows that AGE-induced p-ERK1/2, α-SMA expression, and loss of E-cadherin are regulated by the ERK1/2 MAP kinase pathway. Results show that AGE-induced p-ERK1/2, up-regulation of α-SMA and decrease in E-cadherin expression by NRK52E cells at 24 hours are completely blocked by overexpression of DN-MEK1. However, cells with overexpression of WT-MEK1 show no significant increase in p-ERK1/2 and α-SMA or decrease in E-cadherin expression when compared to empty control vector. Each sample represents the results from at least three independent experiments.

    Journal: The American Journal of Pathology

    Article Title: Advanced Glycation End Products Induce Tubular Epithelial-Myofibroblast Transition through the RAGE-ERK1/2 MAP Kinase Signaling Pathway

    doi:

    Figure Lengend Snippet: Western blot analysis shows that AGE-induced p-ERK1/2, α-SMA expression, and loss of E-cadherin are regulated by the ERK1/2 MAP kinase pathway. Results show that AGE-induced p-ERK1/2, up-regulation of α-SMA and decrease in E-cadherin expression by NRK52E cells at 24 hours are completely blocked by overexpression of DN-MEK1. However, cells with overexpression of WT-MEK1 show no significant increase in p-ERK1/2 and α-SMA or decrease in E-cadherin expression when compared to empty control vector. Each sample represents the results from at least three independent experiments.

    Article Snippet: Specifically, AGEs are able to activate ERK1/2 within 15 minutes and this is blocked by a soluble truncated receptor for AGE, sRAGE, and an inhibitor to ERK1/2, PD98059.

    Techniques: Western Blot, Expressing, Over Expression, Plasmid Preparation

    ERK1 − / − ; ERK2 flox/flox ; Col2a1-Cre embryos. (A) Immunofluorescence using anti-ERK1 and ERK2 antibody showed reduced immunoreactivity in chondrocytes in the tibiae of ERK1 − / − ; ERK2 flox/flox ; Col2a1-Cre embryos at E18.5.

    Journal:

    Article Title: Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿ †

    doi: 10.1128/MCB.01549-08

    Figure Lengend Snippet: ERK1 − / − ; ERK2 flox/flox ; Col2a1-Cre embryos. (A) Immunofluorescence using anti-ERK1 and ERK2 antibody showed reduced immunoreactivity in chondrocytes in the tibiae of ERK1 − / − ; ERK2 flox/flox ; Col2a1-Cre embryos at E18.5.

    Article Snippet: The cartilage phenotype of ERK1 , ERK2 conditional knockout mice was substantially alleviated by one functional allele of either ERK1 or ERK2, indicating that one allele of either ERK1 or ERK2 is sufficient for restoring the growth plate architecture and chondrocyte proliferation.

    Techniques: Immunofluorescence

    Delayed formation of primary ossification centers in ERK1 − / − ; ERK2 flox/flox ; Prx1-Cre mice. (A) Immunohistochemistry for type X collagen showed a widening of the zone of hypertrophic chondrocytes in the tibiae of ERK1 − / −

    Journal:

    Article Title: Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿ †

    doi: 10.1128/MCB.01549-08

    Figure Lengend Snippet: Delayed formation of primary ossification centers in ERK1 − / − ; ERK2 flox/flox ; Prx1-Cre mice. (A) Immunohistochemistry for type X collagen showed a widening of the zone of hypertrophic chondrocytes in the tibiae of ERK1 − / −

    Article Snippet: The cartilage phenotype of ERK1 , ERK2 conditional knockout mice was substantially alleviated by one functional allele of either ERK1 or ERK2, indicating that one allele of either ERK1 or ERK2 is sufficient for restoring the growth plate architecture and chondrocyte proliferation.

    Techniques: Mouse Assay, Immunohistochemistry

    In situ hybridization analyses of the femur (A, B) and calvaria (C), showing normal levels of Col1a1 , Runx2 , and Osterix ( Osx ) expression and markedly decreased Osteocalcin ( OCN ) expression in ERK1 − / − ; ERK2 flox/flox ; Prx1-Cre embryos and

    Journal:

    Article Title: Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿ †

    doi: 10.1128/MCB.01549-08

    Figure Lengend Snippet: In situ hybridization analyses of the femur (A, B) and calvaria (C), showing normal levels of Col1a1 , Runx2 , and Osterix ( Osx ) expression and markedly decreased Osteocalcin ( OCN ) expression in ERK1 − / − ; ERK2 flox/flox ; Prx1-Cre embryos and

    Article Snippet: The cartilage phenotype of ERK1 , ERK2 conditional knockout mice was substantially alleviated by one functional allele of either ERK1 or ERK2, indicating that one allele of either ERK1 or ERK2 is sufficient for restoring the growth plate architecture and chondrocyte proliferation.

    Techniques: In Situ Hybridization, Expressing

    (A) TRAP staining of the femur showing decreased TRAP-positive cells in ERK1 − / − ; ERK2 flox/flox ; Col2a1-Cre embryos at E16.5. (B) ELF97-based fluorescent TRAP staining in combination with immunofluorescence for ERK protein. Spleen cells

    Journal:

    Article Title: Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿ †

    doi: 10.1128/MCB.01549-08

    Figure Lengend Snippet: (A) TRAP staining of the femur showing decreased TRAP-positive cells in ERK1 − / − ; ERK2 flox/flox ; Col2a1-Cre embryos at E16.5. (B) ELF97-based fluorescent TRAP staining in combination with immunofluorescence for ERK protein. Spleen cells

    Article Snippet: The cartilage phenotype of ERK1 , ERK2 conditional knockout mice was substantially alleviated by one functional allele of either ERK1 or ERK2, indicating that one allele of either ERK1 or ERK2 is sufficient for restoring the growth plate architecture and chondrocyte proliferation.

    Techniques: Staining, Immunofluorescence

    ERK1 −/− ; ERK2 flox/flox ; Prx1-Cre embryos and mice. (A) Skeletal preparation after alizarin red and alcian blue staining at P1. Limbs were severely deformed in ERK1 −/− ; ERK2 flox/flox ; Prx1-Cre mice. (B) Skeletal preparation

    Journal:

    Article Title: Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿ †

    doi: 10.1128/MCB.01549-08

    Figure Lengend Snippet: ERK1 −/− ; ERK2 flox/flox ; Prx1-Cre embryos and mice. (A) Skeletal preparation after alizarin red and alcian blue staining at P1. Limbs were severely deformed in ERK1 −/− ; ERK2 flox/flox ; Prx1-Cre mice. (B) Skeletal preparation

    Article Snippet: The cartilage phenotype of ERK1 , ERK2 conditional knockout mice was substantially alleviated by one functional allele of either ERK1 or ERK2, indicating that one allele of either ERK1 or ERK2 is sufficient for restoring the growth plate architecture and chondrocyte proliferation.

    Techniques: Mouse Assay, Staining

    (A) Semiquantitative reverse transcription-PCR showing reduced Osteocalcin ( OCN ) and Bone sialoprotein ( BSP ) expression in primary ERK1 −/− ; ERK2 flox/flox calvarium mesenchymal cells that were infected with Ad expressing Cre recombinase

    Journal:

    Article Title: Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿ †

    doi: 10.1128/MCB.01549-08

    Figure Lengend Snippet: (A) Semiquantitative reverse transcription-PCR showing reduced Osteocalcin ( OCN ) and Bone sialoprotein ( BSP ) expression in primary ERK1 −/− ; ERK2 flox/flox calvarium mesenchymal cells that were infected with Ad expressing Cre recombinase

    Article Snippet: The cartilage phenotype of ERK1 , ERK2 conditional knockout mice was substantially alleviated by one functional allele of either ERK1 or ERK2, indicating that one allele of either ERK1 or ERK2 is sufficient for restoring the growth plate architecture and chondrocyte proliferation.

    Techniques: Polymerase Chain Reaction, Expressing, Infection

    Ectopic cartilage formation in the perichondria of ERK1 − / − ; ERK2 flox/flox ; Prx1-Cre embryos. (A) Alcian blue staining and in situ hybridization of the femur at E15.5. The ectopic cartilage (arrowheads) in the perichondrium expresses Sox9

    Journal:

    Article Title: Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿Extracellular Signal-Regulated Kinase 1 (ERK1) and ERK2 Play Essential Roles in Osteoblast Differentiation and in Supporting Osteoclastogenesis ▿ †

    doi: 10.1128/MCB.01549-08

    Figure Lengend Snippet: Ectopic cartilage formation in the perichondria of ERK1 − / − ; ERK2 flox/flox ; Prx1-Cre embryos. (A) Alcian blue staining and in situ hybridization of the femur at E15.5. The ectopic cartilage (arrowheads) in the perichondrium expresses Sox9

    Article Snippet: The cartilage phenotype of ERK1 , ERK2 conditional knockout mice was substantially alleviated by one functional allele of either ERK1 or ERK2, indicating that one allele of either ERK1 or ERK2 is sufficient for restoring the growth plate architecture and chondrocyte proliferation.

    Techniques: Staining, In Situ Hybridization

    Expressions and prognostic roles of MKP-4/ERK1/2 in HCC patients. a Western blot analysis revealed a lower expression of MKP-4 or higher expressions of ERK1/2, p-ERK1/2 in hepatocellular carcinoma (T) and adjacent non-tumorous tissues (N). b The bar chart demonstrates the ratio of MKP-4, ERK1/2 and p-ERK1/2 to GAPDH by quantitative analysis. * P

    Journal: Cancer Cell International

    Article Title: MKP-4 suppresses hepatocarcinogenesis by targeting ERK1/2 pathway

    doi: 10.1186/s12935-019-0776-3

    Figure Lengend Snippet: Expressions and prognostic roles of MKP-4/ERK1/2 in HCC patients. a Western blot analysis revealed a lower expression of MKP-4 or higher expressions of ERK1/2, p-ERK1/2 in hepatocellular carcinoma (T) and adjacent non-tumorous tissues (N). b The bar chart demonstrates the ratio of MKP-4, ERK1/2 and p-ERK1/2 to GAPDH by quantitative analysis. * P

    Article Snippet: After that, they were incubated with anti-MKP-4 (anti-mouse, 1:500; Immunoway, USA) and anti-ERK1/2 (anti-rabbit, 1:500; Santa Cruz Biotechnology) at 4 °C overnight.

    Techniques: Western Blot, Expressing

    Schematic diagram of proposed mechanism. MKP-4 expression may regulate hepatocellular carcinoma cells proliferation and stemness by inhibiting the phosphorylation of ERK1/2 and enhancing expression of CyclinD1 and c-Myc

    Journal: Cancer Cell International

    Article Title: MKP-4 suppresses hepatocarcinogenesis by targeting ERK1/2 pathway

    doi: 10.1186/s12935-019-0776-3

    Figure Lengend Snippet: Schematic diagram of proposed mechanism. MKP-4 expression may regulate hepatocellular carcinoma cells proliferation and stemness by inhibiting the phosphorylation of ERK1/2 and enhancing expression of CyclinD1 and c-Myc

    Article Snippet: After that, they were incubated with anti-MKP-4 (anti-mouse, 1:500; Immunoway, USA) and anti-ERK1/2 (anti-rabbit, 1:500; Santa Cruz Biotechnology) at 4 °C overnight.

    Techniques: Expressing

    MKP-4 interacts with ERK1/2 in liver tumor cells and tissues. a The results of mass spectrometry in HCC tissues. b Verification of the interaction between MKP-4 and ERK1/2 in HCC tissues using immunoprecipitation assay. c Reciprocal immunoprecipitation of MKP-4 and ERK1/2 in HepG2 cells. Lysates of HepG2 cells were immunoprecipitated with anti-MKP-4, anti-ERK1/2 antibodies or control IgG. The immunoprecipitates were subjected to western blot analysis with anti-ERK1/2 and anti-MKP-4 antibodies. d Immunofluorescence analysis of MKP-4 and ERK1/2 in HepG2 cells. HepG2 cells were subjected to immunofluorescence assay using anti-MKP-4 and anti-ERK1/2 antibodies. Scale bar: 50 μm

    Journal: Cancer Cell International

    Article Title: MKP-4 suppresses hepatocarcinogenesis by targeting ERK1/2 pathway

    doi: 10.1186/s12935-019-0776-3

    Figure Lengend Snippet: MKP-4 interacts with ERK1/2 in liver tumor cells and tissues. a The results of mass spectrometry in HCC tissues. b Verification of the interaction between MKP-4 and ERK1/2 in HCC tissues using immunoprecipitation assay. c Reciprocal immunoprecipitation of MKP-4 and ERK1/2 in HepG2 cells. Lysates of HepG2 cells were immunoprecipitated with anti-MKP-4, anti-ERK1/2 antibodies or control IgG. The immunoprecipitates were subjected to western blot analysis with anti-ERK1/2 and anti-MKP-4 antibodies. d Immunofluorescence analysis of MKP-4 and ERK1/2 in HepG2 cells. HepG2 cells were subjected to immunofluorescence assay using anti-MKP-4 and anti-ERK1/2 antibodies. Scale bar: 50 μm

    Article Snippet: After that, they were incubated with anti-MKP-4 (anti-mouse, 1:500; Immunoway, USA) and anti-ERK1/2 (anti-rabbit, 1:500; Santa Cruz Biotechnology) at 4 °C overnight.

    Techniques: Mass Spectrometry, Immunoprecipitation, Western Blot, Immunofluorescence

    MKP-4 regulates the phosphorylation of ERK1/2 in liver tumor cells. a The expressions of MKP-4 in LO2 and different liver tumor cells were detected by using western blot. b We used RNA interference to knockdown MKP-4 expression in HepG2 or SK-Hep1 cells and chose best interfering efficiency. The bar chart demonstrated the ratio of MKP-4 expression to GAPDH by densitometry. The data were mean ± SEM of three independent experiments (* P

    Journal: Cancer Cell International

    Article Title: MKP-4 suppresses hepatocarcinogenesis by targeting ERK1/2 pathway

    doi: 10.1186/s12935-019-0776-3

    Figure Lengend Snippet: MKP-4 regulates the phosphorylation of ERK1/2 in liver tumor cells. a The expressions of MKP-4 in LO2 and different liver tumor cells were detected by using western blot. b We used RNA interference to knockdown MKP-4 expression in HepG2 or SK-Hep1 cells and chose best interfering efficiency. The bar chart demonstrated the ratio of MKP-4 expression to GAPDH by densitometry. The data were mean ± SEM of three independent experiments (* P

    Article Snippet: After that, they were incubated with anti-MKP-4 (anti-mouse, 1:500; Immunoway, USA) and anti-ERK1/2 (anti-rabbit, 1:500; Santa Cruz Biotechnology) at 4 °C overnight.

    Techniques: Western Blot, Expressing

    MKP-4 inhibits the tumorigenicity of HCC by targeting ERK1/2 pathway in vivo. a MKP-4 inhibits tumor growth of HepG2 cells in vivo. Control, MKP-4 silenced or MKP-4 overexpressed HepG2 cells were injected into BALB/c nude mice. b , c Tumor weight and volume harvested from the nude mice in different groups after 4 weeks. * P

    Journal: Cancer Cell International

    Article Title: MKP-4 suppresses hepatocarcinogenesis by targeting ERK1/2 pathway

    doi: 10.1186/s12935-019-0776-3

    Figure Lengend Snippet: MKP-4 inhibits the tumorigenicity of HCC by targeting ERK1/2 pathway in vivo. a MKP-4 inhibits tumor growth of HepG2 cells in vivo. Control, MKP-4 silenced or MKP-4 overexpressed HepG2 cells were injected into BALB/c nude mice. b , c Tumor weight and volume harvested from the nude mice in different groups after 4 weeks. * P

    Article Snippet: After that, they were incubated with anti-MKP-4 (anti-mouse, 1:500; Immunoway, USA) and anti-ERK1/2 (anti-rabbit, 1:500; Santa Cruz Biotechnology) at 4 °C overnight.

    Techniques: In Vivo, Injection, Mouse Assay

    MKP-4 inhibits cell proliferation and cancer stem cell (CSC) traits through ERK1/2 pathway. a MKP-4 reduced the colony formation of HepG2 or SK-Hep1 cells via the interaction with ERK1/2. For colony formation assay, MKP-4 knockdown, MKP-4 overexpression or pre-incubation with 10 μM PD98059 by 24 h cells were seeded into each well of six-well-plate colonies and stained with crystal violet after 2 weeks. b CCK-8 assay showed that overexpression of MKP-4 or pre-incubation with 10 μM PD98059 by 24 h inhibited cell proliferation in HepG2 and SK-Hep1 cells while MKP-4 depletion promoted cell proliferation. The data are mean ± SEM of three independent experiments. (* P

    Journal: Cancer Cell International

    Article Title: MKP-4 suppresses hepatocarcinogenesis by targeting ERK1/2 pathway

    doi: 10.1186/s12935-019-0776-3

    Figure Lengend Snippet: MKP-4 inhibits cell proliferation and cancer stem cell (CSC) traits through ERK1/2 pathway. a MKP-4 reduced the colony formation of HepG2 or SK-Hep1 cells via the interaction with ERK1/2. For colony formation assay, MKP-4 knockdown, MKP-4 overexpression or pre-incubation with 10 μM PD98059 by 24 h cells were seeded into each well of six-well-plate colonies and stained with crystal violet after 2 weeks. b CCK-8 assay showed that overexpression of MKP-4 or pre-incubation with 10 μM PD98059 by 24 h inhibited cell proliferation in HepG2 and SK-Hep1 cells while MKP-4 depletion promoted cell proliferation. The data are mean ± SEM of three independent experiments. (* P

    Article Snippet: After that, they were incubated with anti-MKP-4 (anti-mouse, 1:500; Immunoway, USA) and anti-ERK1/2 (anti-rabbit, 1:500; Santa Cruz Biotechnology) at 4 °C overnight.

    Techniques: Colony Assay, Over Expression, Incubation, Staining, CCK-8 Assay

    Effects of ApoSOD1 on Ca 2+ /Akt/ERK1/2 signaling pathway in NSC-34 motor neurons. ( a ) (Top) Western blotting of SOD1 expression after inactivation with H 2 O 2 (200 mM) for several time points. (Bottom) Representative curve of the time-dependent inactivation of SOD1 in H 2 O 2 . ( b ) Bar graph depicting DCF-DA-detected ROS production in NSC-34 motor neurons exposed to chemical hypoxia in the absence and in the presence of SOD1 (400 ng/ml) or ApoSOD1 (400 ng/ml). Data are expressed as mean±S.E. of three different experiments. * P

    Journal: Cell Death and Differentiation

    Article Title: ApoSOD1 lacking dismutase activity neuroprotects motor neurons exposed to beta-methylamino-L-alanine through the Ca2+/Akt/ERK1/2 prosurvival pathway

    doi: 10.1038/cdd.2016.154

    Figure Lengend Snippet: Effects of ApoSOD1 on Ca 2+ /Akt/ERK1/2 signaling pathway in NSC-34 motor neurons. ( a ) (Top) Western blotting of SOD1 expression after inactivation with H 2 O 2 (200 mM) for several time points. (Bottom) Representative curve of the time-dependent inactivation of SOD1 in H 2 O 2 . ( b ) Bar graph depicting DCF-DA-detected ROS production in NSC-34 motor neurons exposed to chemical hypoxia in the absence and in the presence of SOD1 (400 ng/ml) or ApoSOD1 (400 ng/ml). Data are expressed as mean±S.E. of three different experiments. * P

    Article Snippet: Membranes were blocked with 5% non-fat dry milk in 0.1% Tween 20 (Sigma-Aldrich; 2 mM Tris-HCl and 50 mM NaCl, pH 7.5) for 2 h at room temperature; then membranes were incubated overnight at 4 °C in blocking buffer containing monoclonal antibodies (1:1000) against p-Akt (Cell Signaling Technology Inc.) and p-ERK1/2 (Santa Cruz Biotecnology, Inc.), rabbit polyclonal antibodies (1:1000) against GRP78 (Cell Signaling Technology Inc.) or rabbit polyclonal antibodies (1 : 1000) against caspase-12 (Cell Signaling Technology Inc.).

    Techniques: Western Blot, Expressing

    Effects of SOD1 on [Ca 2+ ] i and ERK1/2 and Akt phosphorylation in NSC-34 motor neurons. ( a and b ) (Top) Representative images of NSC-34 motor neurons loaded with Fura-2/AM in control conditions. (Bottom a ) Single-cell trace depicting [Ca 2+ ] i before and after SOD1 administration. (Bottom b ) Quantification of the effect of SOD1, SOD1 in Ca 2+ -free and SOD1 in Ca 2+ -free + thapsigargin (Tg) on [Ca 2+ ] i SOD1 significantly increases [Ca 2+ ] i . Data are expressed as mean±S.E. of three different experiments performed on 30/50 cells. * P

    Journal: Cell Death and Differentiation

    Article Title: ApoSOD1 lacking dismutase activity neuroprotects motor neurons exposed to beta-methylamino-L-alanine through the Ca2+/Akt/ERK1/2 prosurvival pathway

    doi: 10.1038/cdd.2016.154

    Figure Lengend Snippet: Effects of SOD1 on [Ca 2+ ] i and ERK1/2 and Akt phosphorylation in NSC-34 motor neurons. ( a and b ) (Top) Representative images of NSC-34 motor neurons loaded with Fura-2/AM in control conditions. (Bottom a ) Single-cell trace depicting [Ca 2+ ] i before and after SOD1 administration. (Bottom b ) Quantification of the effect of SOD1, SOD1 in Ca 2+ -free and SOD1 in Ca 2+ -free + thapsigargin (Tg) on [Ca 2+ ] i SOD1 significantly increases [Ca 2+ ] i . Data are expressed as mean±S.E. of three different experiments performed on 30/50 cells. * P

    Article Snippet: Membranes were blocked with 5% non-fat dry milk in 0.1% Tween 20 (Sigma-Aldrich; 2 mM Tris-HCl and 50 mM NaCl, pH 7.5) for 2 h at room temperature; then membranes were incubated overnight at 4 °C in blocking buffer containing monoclonal antibodies (1:1000) against p-Akt (Cell Signaling Technology Inc.) and p-ERK1/2 (Santa Cruz Biotecnology, Inc.), rabbit polyclonal antibodies (1:1000) against GRP78 (Cell Signaling Technology Inc.) or rabbit polyclonal antibodies (1 : 1000) against caspase-12 (Cell Signaling Technology Inc.).

    Techniques:

    Effects of ApoSOD1, SOD1, rSOD1 wt and rSOD1 G93A on Ca 2+ /Akt/ERK1/2 signaling pathway in rat primary motor neurons. ( a ) Immunocytochemical images of SMI-32 and Hoechst. ( b – f ) Representative single-cell traces ( b – e ) and quantification ( f ) of the effect on [Ca 2+ ] i of SOD1 ( b ), ApoSOD1 ( c ), rSOD1 wt ( d ) and rSOD1 G93A ( e ), all at 400 ng/ml in normal Krebs solution. Data are expressed as mean±S.E. of three different experiments performed on 30/50 cells for each group. * P

    Journal: Cell Death and Differentiation

    Article Title: ApoSOD1 lacking dismutase activity neuroprotects motor neurons exposed to beta-methylamino-L-alanine through the Ca2+/Akt/ERK1/2 prosurvival pathway

    doi: 10.1038/cdd.2016.154

    Figure Lengend Snippet: Effects of ApoSOD1, SOD1, rSOD1 wt and rSOD1 G93A on Ca 2+ /Akt/ERK1/2 signaling pathway in rat primary motor neurons. ( a ) Immunocytochemical images of SMI-32 and Hoechst. ( b – f ) Representative single-cell traces ( b – e ) and quantification ( f ) of the effect on [Ca 2+ ] i of SOD1 ( b ), ApoSOD1 ( c ), rSOD1 wt ( d ) and rSOD1 G93A ( e ), all at 400 ng/ml in normal Krebs solution. Data are expressed as mean±S.E. of three different experiments performed on 30/50 cells for each group. * P

    Article Snippet: Membranes were blocked with 5% non-fat dry milk in 0.1% Tween 20 (Sigma-Aldrich; 2 mM Tris-HCl and 50 mM NaCl, pH 7.5) for 2 h at room temperature; then membranes were incubated overnight at 4 °C in blocking buffer containing monoclonal antibodies (1:1000) against p-Akt (Cell Signaling Technology Inc.) and p-ERK1/2 (Santa Cruz Biotecnology, Inc.), rabbit polyclonal antibodies (1:1000) against GRP78 (Cell Signaling Technology Inc.) or rabbit polyclonal antibodies (1 : 1000) against caspase-12 (Cell Signaling Technology Inc.).

    Techniques:

    Effects of the inhibition of ERK1/2/Akt signaling pathway on SOD1-induced neuroprotection in NSC-34 motor neurons exposed to L-BMAA. ( a ) Western blotting and quantification of p-GSK3 β and GSK3 β expression in NSC-34 cells transfected with Akt D− (2 μ g/ μ l) and exposed to SOD1 (400 ng/ml/10 min). NSC-34 cells were transfected with Akt D− or Mock 48 h before the exposure to SOD1. The transfection medium containing vectors was incubated for 5 h and then replaced with fresh medium. Data are expressed as mean±S.E. of three different experimental sessions. * P

    Journal: Cell Death and Differentiation

    Article Title: ApoSOD1 lacking dismutase activity neuroprotects motor neurons exposed to beta-methylamino-L-alanine through the Ca2+/Akt/ERK1/2 prosurvival pathway

    doi: 10.1038/cdd.2016.154

    Figure Lengend Snippet: Effects of the inhibition of ERK1/2/Akt signaling pathway on SOD1-induced neuroprotection in NSC-34 motor neurons exposed to L-BMAA. ( a ) Western blotting and quantification of p-GSK3 β and GSK3 β expression in NSC-34 cells transfected with Akt D− (2 μ g/ μ l) and exposed to SOD1 (400 ng/ml/10 min). NSC-34 cells were transfected with Akt D− or Mock 48 h before the exposure to SOD1. The transfection medium containing vectors was incubated for 5 h and then replaced with fresh medium. Data are expressed as mean±S.E. of three different experimental sessions. * P

    Article Snippet: Membranes were blocked with 5% non-fat dry milk in 0.1% Tween 20 (Sigma-Aldrich; 2 mM Tris-HCl and 50 mM NaCl, pH 7.5) for 2 h at room temperature; then membranes were incubated overnight at 4 °C in blocking buffer containing monoclonal antibodies (1:1000) against p-Akt (Cell Signaling Technology Inc.) and p-ERK1/2 (Santa Cruz Biotecnology, Inc.), rabbit polyclonal antibodies (1:1000) against GRP78 (Cell Signaling Technology Inc.) or rabbit polyclonal antibodies (1 : 1000) against caspase-12 (Cell Signaling Technology Inc.).

    Techniques: Inhibition, Western Blot, Expressing, Transfection, Incubation

    Effects of the recombinant SOD1 G93A on Akt/ERK1/2 signaling pathway and on cell survival in NSC-34 motor neurons exposed to L-BMAA. ( a ) Representative western blotting and quantification of the effect of SOD1, rSOD1 wt and rSOD1 G93A (400 ng/ml/10 min) on p-ERK1/2 and ERK1/2 expression. ( b ) Representative western blotting and quantification of the effect of SOD1, rSOD1 wt and rSOD1 G93A (400 ng/ml/10 min) on p-Akt and Akt1/2/3 expression. ( a and b ) Data are expressed as mean±SE of three different experimental sessions. * P

    Journal: Cell Death and Differentiation

    Article Title: ApoSOD1 lacking dismutase activity neuroprotects motor neurons exposed to beta-methylamino-L-alanine through the Ca2+/Akt/ERK1/2 prosurvival pathway

    doi: 10.1038/cdd.2016.154

    Figure Lengend Snippet: Effects of the recombinant SOD1 G93A on Akt/ERK1/2 signaling pathway and on cell survival in NSC-34 motor neurons exposed to L-BMAA. ( a ) Representative western blotting and quantification of the effect of SOD1, rSOD1 wt and rSOD1 G93A (400 ng/ml/10 min) on p-ERK1/2 and ERK1/2 expression. ( b ) Representative western blotting and quantification of the effect of SOD1, rSOD1 wt and rSOD1 G93A (400 ng/ml/10 min) on p-Akt and Akt1/2/3 expression. ( a and b ) Data are expressed as mean±SE of three different experimental sessions. * P

    Article Snippet: Membranes were blocked with 5% non-fat dry milk in 0.1% Tween 20 (Sigma-Aldrich; 2 mM Tris-HCl and 50 mM NaCl, pH 7.5) for 2 h at room temperature; then membranes were incubated overnight at 4 °C in blocking buffer containing monoclonal antibodies (1:1000) against p-Akt (Cell Signaling Technology Inc.) and p-ERK1/2 (Santa Cruz Biotecnology, Inc.), rabbit polyclonal antibodies (1:1000) against GRP78 (Cell Signaling Technology Inc.) or rabbit polyclonal antibodies (1 : 1000) against caspase-12 (Cell Signaling Technology Inc.).

    Techniques: Recombinant, Western Blot, Expressing