phospho p38 mapk thr180 tyr182 rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 rabbit mab
    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Phospho P38 Mapk Thr180 Tyr182 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 mapk thr180 tyr182 rabbit mab - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model"

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    Journal: Neural Regeneration Research

    doi: 10.4103/1673-5374.391193

    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: p38 MAPK inhibitor.
    Figure Legend Snippet: Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: p38 MAPK inhibitor.

    Techniques Used: In Vivo, In Vitro

    p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced cell death. (A) The effect of different concentrations of glutamate on R28 cell viability over 24 hours ( n = 5). (B) Light microscopy was used to examine the effect of SB202190 on 10 mM glutamate-treated R28 cells. (C) Effect of SB202190 on the cell viability of R28 cells treated with 10 mM glutamate for 24 hours ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . control group (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. (D) The effect of SB202190 on the morphology of R28 cells was observed under a light microscope. Arrows indicate dead cells. Scale bars: 20 µm.
    Figure Legend Snippet: p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced cell death. (A) The effect of different concentrations of glutamate on R28 cell viability over 24 hours ( n = 5). (B) Light microscopy was used to examine the effect of SB202190 on 10 mM glutamate-treated R28 cells. (C) Effect of SB202190 on the cell viability of R28 cells treated with 10 mM glutamate for 24 hours ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . control group (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. (D) The effect of SB202190 on the morphology of R28 cells was observed under a light microscope. Arrows indicate dead cells. Scale bars: 20 µm.

    Techniques Used: Light Microscopy, Comparison

    p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced ferroptosis. (A) Propidium iodide (PI; red) and Hoechst 33342 (blue) staining of R28 cells treated with glutamate, SB202190 and ferrostatin-1 as indicated for 24 hours. Scale bar: 100 µm. (B) Quantification of the results shown in A ( n = 5). (C) Transmission electron microscope images of R28 cells treated with glutamate and SB202190. Scale bar: 1 µm. (D) R28 cells treated as indicated were stained with the fluorescent probe Mito -FerroOrange to determine ferrous ion levels. FerroOrange undergoes an irreversible reaction with intracellular Fe 2+ in live cells, resulting in orange fluorescence emission. Ex: 543 nm and Em: 580 nm. Scale bar: 50 µm. (E) Quantification of mean density of FerroOrange fluorescence in the groups ( n = 5). (F) R28 cells treated as indicated were stained with Liperfluo for measurement of lipid peroxides. Liperfluo is selectively oxidized by lipid peroxides, and the oxidized form of Liperfluo exhibits strong green fluorescence at the cell membrane. Scale bar: 50 µm. (G) Comparison of mean density of Liperfluor fluorescence in the groups ( n = 5). (H, I) Malondialdehyde (MDA; H) and glutathione (GSH; I) levels in R28 cells after glutamate and SB202190 treatment ( n = 3). ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.
    Figure Legend Snippet: p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced ferroptosis. (A) Propidium iodide (PI; red) and Hoechst 33342 (blue) staining of R28 cells treated with glutamate, SB202190 and ferrostatin-1 as indicated for 24 hours. Scale bar: 100 µm. (B) Quantification of the results shown in A ( n = 5). (C) Transmission electron microscope images of R28 cells treated with glutamate and SB202190. Scale bar: 1 µm. (D) R28 cells treated as indicated were stained with the fluorescent probe Mito -FerroOrange to determine ferrous ion levels. FerroOrange undergoes an irreversible reaction with intracellular Fe 2+ in live cells, resulting in orange fluorescence emission. Ex: 543 nm and Em: 580 nm. Scale bar: 50 µm. (E) Quantification of mean density of FerroOrange fluorescence in the groups ( n = 5). (F) R28 cells treated as indicated were stained with Liperfluo for measurement of lipid peroxides. Liperfluo is selectively oxidized by lipid peroxides, and the oxidized form of Liperfluo exhibits strong green fluorescence at the cell membrane. Scale bar: 50 µm. (G) Comparison of mean density of Liperfluor fluorescence in the groups ( n = 5). (H, I) Malondialdehyde (MDA; H) and glutathione (GSH; I) levels in R28 cells after glutamate and SB202190 treatment ( n = 3). ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.

    Techniques Used: Staining, Transmission Assay, Microscopy, Fluorescence, Membrane, Comparison

    p38 MAPK inhibitor SB202190 regulates ferroptosis in a glutamate R28 cell model by regulating FTL and SAT1. (A) p38 MAPK, p-p38 MAPK, FTL and SAT1 protein expression in R28 cells treated with glutamate and SB202190 for 24 hours ( n = 3). (B–D) Quantification analysis of p-p38 MAPK/ p38 MAPK, FTL and SAT1 protein expression in R28 cells (n = 3). (E–I) Three days after intravitreal injection of NMDA and SB202190, paraffin sections were collected and processed for immunofluorescence experiments to measure the fluorescence intensity of p-p38 MAPK (E, F), FTL (G, H), and SAT1 (I, J) of the retinal sections in each group under a fluorescence microscope ( n = 3). Scale bars: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were repeated. FTL: Ferritin light chain; MAPK: mitogen activated protein kinases.
    Figure Legend Snippet: p38 MAPK inhibitor SB202190 regulates ferroptosis in a glutamate R28 cell model by regulating FTL and SAT1. (A) p38 MAPK, p-p38 MAPK, FTL and SAT1 protein expression in R28 cells treated with glutamate and SB202190 for 24 hours ( n = 3). (B–D) Quantification analysis of p-p38 MAPK/ p38 MAPK, FTL and SAT1 protein expression in R28 cells (n = 3). (E–I) Three days after intravitreal injection of NMDA and SB202190, paraffin sections were collected and processed for immunofluorescence experiments to measure the fluorescence intensity of p-p38 MAPK (E, F), FTL (G, H), and SAT1 (I, J) of the retinal sections in each group under a fluorescence microscope ( n = 3). Scale bars: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were repeated. FTL: Ferritin light chain; MAPK: mitogen activated protein kinases.

    Techniques Used: Expressing, Injection, Immunofluorescence, Fluorescence, Microscopy, Comparison

    p38 MAPK inhibitor SB202190 alleviates lipid peroxidation by regulating the SLC7A11/GSH/GPx4 pathway. (A, B) Flow cytometry was used to detect the effect of SB202190 on lipid reactive oxygen species production in R28 cells treated as indicated for 24 hours ( n = 4). (C–E) SLC7A11 and GPx4 protein expressions in R28 cells were detected by western blotting ( n = 3). (F–I) Fluorescence microscopy was performed to evaluate the fluorescence intensity changes of and GPx4 (F, G) and SLC7A11 (H, I) in rat retinal sections after intraluminal injection of NMDA and SB202190 for 3 days ( n = 3). Scale bars in F and H: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. GPx4: Glutathione peroxidase 4; GSH: glutathione; MAPK: mitogen-activated protein kinase.
    Figure Legend Snippet: p38 MAPK inhibitor SB202190 alleviates lipid peroxidation by regulating the SLC7A11/GSH/GPx4 pathway. (A, B) Flow cytometry was used to detect the effect of SB202190 on lipid reactive oxygen species production in R28 cells treated as indicated for 24 hours ( n = 4). (C–E) SLC7A11 and GPx4 protein expressions in R28 cells were detected by western blotting ( n = 3). (F–I) Fluorescence microscopy was performed to evaluate the fluorescence intensity changes of and GPx4 (F, G) and SLC7A11 (H, I) in rat retinal sections after intraluminal injection of NMDA and SB202190 for 3 days ( n = 3). Scale bars in F and H: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. GPx4: Glutathione peroxidase 4; GSH: glutathione; MAPK: mitogen-activated protein kinase.

    Techniques Used: Flow Cytometry, Western Blot, Fluorescence, Microscopy, Injection, Comparison

    p38 MAPK inhibitor SB202190 attenuates N-methyl-D-aspartate (NMDA)-induced damage in the rat retina. (A, B) Qualitative observation (A) and quantitative analysis (B) of the effects of SB202190 on NMDA-induced retinal ganglion cell (RGC) injury in rat retina ( n = 5). Three days after intravitreal injection of NMDA and SB202190, retinal plating was performed. RGCs were fluorescently labeled with Brn3a antibody and surviving RGC exhibited strong green fluorescence under a fluorescence microscope. Scale bar: 50 μm. (C) Effects of SB202190 on retinal morphology in NMDA model rats. Hematoxylin and eosin staining was performed 3 days after intravitreal injection of NMDA and SB202190. Scale bar: 50 µm. (D) Effect of SB202190 on RGC density (cells/100 µm) in NMDA model rats 3 days after intravitreal injection ( n = 4). (E, F) Thickness of the retinal ganglion cell body complex (GCC) at 500, 1000, 1500, and 2000 µm from the optic disc 3 days after intravitreal injection of NMDA and SB202190. (G–I) Flash visual evoked potentials of rats 3 days after intravitreal injection of NMDA and SB202190 ( n = 4). GCL: Ganglion cell layer; INL: inner nuclear layer; IPL: inner plexiform layer; ONL: outer nuclear layer; RGC: retinal ganglion cell. Inner retina consists of GCL and IPL. ** P < 0.01, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.
    Figure Legend Snippet: p38 MAPK inhibitor SB202190 attenuates N-methyl-D-aspartate (NMDA)-induced damage in the rat retina. (A, B) Qualitative observation (A) and quantitative analysis (B) of the effects of SB202190 on NMDA-induced retinal ganglion cell (RGC) injury in rat retina ( n = 5). Three days after intravitreal injection of NMDA and SB202190, retinal plating was performed. RGCs were fluorescently labeled with Brn3a antibody and surviving RGC exhibited strong green fluorescence under a fluorescence microscope. Scale bar: 50 μm. (C) Effects of SB202190 on retinal morphology in NMDA model rats. Hematoxylin and eosin staining was performed 3 days after intravitreal injection of NMDA and SB202190. Scale bar: 50 µm. (D) Effect of SB202190 on RGC density (cells/100 µm) in NMDA model rats 3 days after intravitreal injection ( n = 4). (E, F) Thickness of the retinal ganglion cell body complex (GCC) at 500, 1000, 1500, and 2000 µm from the optic disc 3 days after intravitreal injection of NMDA and SB202190. (G–I) Flash visual evoked potentials of rats 3 days after intravitreal injection of NMDA and SB202190 ( n = 4). GCL: Ganglion cell layer; INL: inner nuclear layer; IPL: inner plexiform layer; ONL: outer nuclear layer; RGC: retinal ganglion cell. Inner retina consists of GCL and IPL. ** P < 0.01, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.

    Techniques Used: Injection, Labeling, Fluorescence, Microscopy, Staining, Comparison

    A schematic model for the mechanism of p38 MAPK inhibitor SB202190 in alleviating ferroptosis in the glutamate-induced retinal excitotoxic glaucoma model. The p38 MAPK inhibitor sB202190 inhibits ferroptosis in the glutamatergic excitotoxicity glaucoma model by upregulation of ferritin light chain (FTL), downregulation of SAT1 and regulation of the SLC7A11/GSH/GPx4 signaling pathway. GPx4: Glutathione peroxidase 4; GSH: glutathione; GSSG: glutathione disulfide; MAPK: mitogen-activated protein kinase; ROS: reactive oxygen species.
    Figure Legend Snippet: A schematic model for the mechanism of p38 MAPK inhibitor SB202190 in alleviating ferroptosis in the glutamate-induced retinal excitotoxic glaucoma model. The p38 MAPK inhibitor sB202190 inhibits ferroptosis in the glutamatergic excitotoxicity glaucoma model by upregulation of ferritin light chain (FTL), downregulation of SAT1 and regulation of the SLC7A11/GSH/GPx4 signaling pathway. GPx4: Glutathione peroxidase 4; GSH: glutathione; GSSG: glutathione disulfide; MAPK: mitogen-activated protein kinase; ROS: reactive oxygen species.

    Techniques Used:

    phospho p44 42 mapk erk1 2 thr202 tyr204 xp rabbit mab  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc phospho p44 42 mapk erk1 2 thr202 tyr204 xp rabbit mab
    Phospho P44 42 Mapk Erk1 2 Thr202 Tyr204 Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit monoclonal anti phospho p38 mapk thr180 tyr182 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti phospho p38 mapk thr180 tyr182 antibody
    Rabbit Monoclonal Anti Phospho P38 Mapk Thr180 Tyr182 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit monoclonal anti phospho p44 42 mapk erk1 2  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti phospho p44 42 mapk erk1 2
    a , b Volcano plot of microRNA analyzes of human samples, separated by sex for mature ( a ) and hairpin ( b ) microRNAs. We used DESEq2 for DE analysis and Benjamini-Hochberg for multiple test correction. The x -axis shows the log2 fold change in ALS vs CTR, whereas the y -axis shows the -log10 p-value. Orange and blue dots represent DEGs in females and males, respectively. miRNA-mediated regulation in ALS reveals a notable sex-dependent pattern. Male ALS patients exhibit a more pronounced downregulation of mature and hairpin miRNAs compared to females. c Heatmap showing mature miRNA expression changes for the identified ALS clusters. miRNA candidates significant in at least one condition were considered (p < 0.05). The scale (left-hand side) shows the range of log2 fold change values. d Volcano plot of differentially expressed proteins (DEP) in human samples (calculated with limma, two-sided test; multiple test correction with Benjamini-Hochberg). The x -axis shows the log2 fold change ALS and CTR, whereas the y -axis shows the -log10 p-values. Orange and blue dots represent DEGs in females and males, respectively. 379 DEPs in males and 251 in females. ANXA2 emerges as the sole protein downregulated in both sexes (p < 0.1). e , f Functional enrichment and unsupervised clustering using REVIGO, a tool used for summarizing Gene Ontology (GO) terms. Important pathway clusters are revealed in both sexes, including synaptic function, immune response, and ECM/cytoskeleton. Females ( e ) exhibited enrichment in transmembrane transport, lipid metabolism, development, catalytic activity, <t>and</t> <t>ERK1/2</t> signaling, while males ( f ) showed enrichment in cell metabolism and tyrosine kinase-related pathways. The size of the circles represents the number of genes in the GO-BP terms, and the color of each node represents the enrichment FDR values. g , h Uniform manifold approximation and projection (UMAP) of MOFA factor analysis for sex ( g ) and condition (ALS vs. CTR) ( h ). Sex ( g ) emerges as a robust differentiating factor when integrating transcriptomic, small RNA, and proteomic data. i MOFA correlation analysis displays which factors are dominated by which omic layer. Components of the representative factors 3 (males) and 12 (females) are displayed in feature-weight plots (bottom part). These factors are the ones that better correlate with disease conditions for each sex.
    Rabbit Monoclonal Anti Phospho P44 42 Mapk Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti phospho p44 42 mapk erk1 2/product/Cell Signaling Technology Inc
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    1) Product Images from "Multiomic ALS signatures highlight subclusters and sex differences suggesting the MAPK pathway as therapeutic target"

    Article Title: Multiomic ALS signatures highlight subclusters and sex differences suggesting the MAPK pathway as therapeutic target

    Journal: Nature Communications

    doi: 10.1038/s41467-024-49196-y

    a , b Volcano plot of microRNA analyzes of human samples, separated by sex for mature ( a ) and hairpin ( b ) microRNAs. We used DESEq2 for DE analysis and Benjamini-Hochberg for multiple test correction. The x -axis shows the log2 fold change in ALS vs CTR, whereas the y -axis shows the -log10 p-value. Orange and blue dots represent DEGs in females and males, respectively. miRNA-mediated regulation in ALS reveals a notable sex-dependent pattern. Male ALS patients exhibit a more pronounced downregulation of mature and hairpin miRNAs compared to females. c Heatmap showing mature miRNA expression changes for the identified ALS clusters. miRNA candidates significant in at least one condition were considered (p < 0.05). The scale (left-hand side) shows the range of log2 fold change values. d Volcano plot of differentially expressed proteins (DEP) in human samples (calculated with limma, two-sided test; multiple test correction with Benjamini-Hochberg). The x -axis shows the log2 fold change ALS and CTR, whereas the y -axis shows the -log10 p-values. Orange and blue dots represent DEGs in females and males, respectively. 379 DEPs in males and 251 in females. ANXA2 emerges as the sole protein downregulated in both sexes (p < 0.1). e , f Functional enrichment and unsupervised clustering using REVIGO, a tool used for summarizing Gene Ontology (GO) terms. Important pathway clusters are revealed in both sexes, including synaptic function, immune response, and ECM/cytoskeleton. Females ( e ) exhibited enrichment in transmembrane transport, lipid metabolism, development, catalytic activity, and ERK1/2 signaling, while males ( f ) showed enrichment in cell metabolism and tyrosine kinase-related pathways. The size of the circles represents the number of genes in the GO-BP terms, and the color of each node represents the enrichment FDR values. g , h Uniform manifold approximation and projection (UMAP) of MOFA factor analysis for sex ( g ) and condition (ALS vs. CTR) ( h ). Sex ( g ) emerges as a robust differentiating factor when integrating transcriptomic, small RNA, and proteomic data. i MOFA correlation analysis displays which factors are dominated by which omic layer. Components of the representative factors 3 (males) and 12 (females) are displayed in feature-weight plots (bottom part). These factors are the ones that better correlate with disease conditions for each sex.
    Figure Legend Snippet: a , b Volcano plot of microRNA analyzes of human samples, separated by sex for mature ( a ) and hairpin ( b ) microRNAs. We used DESEq2 for DE analysis and Benjamini-Hochberg for multiple test correction. The x -axis shows the log2 fold change in ALS vs CTR, whereas the y -axis shows the -log10 p-value. Orange and blue dots represent DEGs in females and males, respectively. miRNA-mediated regulation in ALS reveals a notable sex-dependent pattern. Male ALS patients exhibit a more pronounced downregulation of mature and hairpin miRNAs compared to females. c Heatmap showing mature miRNA expression changes for the identified ALS clusters. miRNA candidates significant in at least one condition were considered (p < 0.05). The scale (left-hand side) shows the range of log2 fold change values. d Volcano plot of differentially expressed proteins (DEP) in human samples (calculated with limma, two-sided test; multiple test correction with Benjamini-Hochberg). The x -axis shows the log2 fold change ALS and CTR, whereas the y -axis shows the -log10 p-values. Orange and blue dots represent DEGs in females and males, respectively. 379 DEPs in males and 251 in females. ANXA2 emerges as the sole protein downregulated in both sexes (p < 0.1). e , f Functional enrichment and unsupervised clustering using REVIGO, a tool used for summarizing Gene Ontology (GO) terms. Important pathway clusters are revealed in both sexes, including synaptic function, immune response, and ECM/cytoskeleton. Females ( e ) exhibited enrichment in transmembrane transport, lipid metabolism, development, catalytic activity, and ERK1/2 signaling, while males ( f ) showed enrichment in cell metabolism and tyrosine kinase-related pathways. The size of the circles represents the number of genes in the GO-BP terms, and the color of each node represents the enrichment FDR values. g , h Uniform manifold approximation and projection (UMAP) of MOFA factor analysis for sex ( g ) and condition (ALS vs. CTR) ( h ). Sex ( g ) emerges as a robust differentiating factor when integrating transcriptomic, small RNA, and proteomic data. i MOFA correlation analysis displays which factors are dominated by which omic layer. Components of the representative factors 3 (males) and 12 (females) are displayed in feature-weight plots (bottom part). These factors are the ones that better correlate with disease conditions for each sex.

    Techniques Used: Expressing, Functional Assay, Activity Assay

    rabbit monoclonal anti phospho p44 42 mapk erk1 2  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc rabbit monoclonal anti phospho p44 42 mapk erk1 2

    Rabbit Monoclonal Anti Phospho P44 42 Mapk Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Analyzing efficiency of a lentiviral shRNA knockdown system in human enteroids using western blot and flow cytometry"

    Article Title: Analyzing efficiency of a lentiviral shRNA knockdown system in human enteroids using western blot and flow cytometry

    Journal: STAR Protocols

    doi: 10.1016/j.xpro.2024.103082


    Figure Legend Snippet:

    Techniques Used: Recombinant, Membrane, Staining, Protease Inhibitor, Lysis, Extraction, Bicinchoninic Acid Protein Assay, Transduction, Positive Control, shRNA, Sequencing, Software, Flow Cytometry, Sterility, Transferring, Adhesive, Aerosol, Cell Counting, Electrophoresis, Microscopy, Purification

    rabbit monoclonal anti phospho p44 42 mapk  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal anti phospho p44 42 mapk

    Rabbit Monoclonal Anti Phospho P44 42 Mapk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti phospho p44 42 mapk/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    1) Product Images from "Oxygen tension-dependent variability in the cancer cell kinome impacts signaling pathways and response to targeted therapies"

    Article Title: Oxygen tension-dependent variability in the cancer cell kinome impacts signaling pathways and response to targeted therapies

    Journal: iScience

    doi: 10.1016/j.isci.2024.110068


    Figure Legend Snippet:

    Techniques Used: Recombinant, Gentle, Lysis, Protease Inhibitor, BrdU Cell Proliferation Assay, Caspase-3 Assay, RNA Sequencing Assay, Mass Spectrometry, Transgenic Assay, TaqMan Assay, Software, Modification, Flow Cytometry, Real-time Polymerase Chain Reaction, Isolation

    anti phospho p44 42 mapk erk1 2 thr202 tyr204 rabbit mab  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc anti phospho p44 42 mapk erk1 2 thr202 tyr204 rabbit mab
    A: Immunoblot analysis of MAPK and PI3K/AKT signaling in the presence of increasing doses of erdafitinib, growth factors- In untreated <t>fibroblasts,</t> <t>ERK1/2</t> and AKT (both pAKT-T308 and pAKT-S473) are activated (in NC, normal control (no DMSO), 0 nM erdafitinib but with DMSO), Erdafitinib treatment at 100 nM dramatically reduce activated pERK1/2 and pAKT with no further reductions detected when fibroblasts were treated with 200 nM or 300 nM erdafitinib (3A-a) . Erdafitinib-inhibited pERK1/2 was not re-activated by the co-addition of IGF-I ( 3A-b ), FGF2 ( 3A-c ) or IGF-2 ( 3A-d ). AKT signaling pathway (pAKT-T308 and pAKT-S473) were consistently activated by IGF-I and IGF-2, but not by FGF2 ( 3A ), at all concentrations of erdafitinib tested (100 nM–300 nM) (see -uncropped.) .B: immunoblot analysis of signaling response in the presence of erdafitinib and growth factors over time- Fibroblasts were co-treated with 200nM erdafitinib and growth factors (IGF-1, IGF-2, FGF-2) for 0.5, 2, 24, 48 hours. Only with IGF-I treatment, activation of AKT was sustained for 48 hours. our data suggest, in our patient fibroblasts, that IGF-I activation of AKT signaling is sustained and independent of the FGFR signaling inhibitory effects of erdafitinib. (see -uncropped.).
    Anti Phospho P44 42 Mapk Erk1 2 Thr202 Tyr204 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Skeletal overgrowth in a pre-pubescent child treated with pan-FGFR inhibitor"

    Article Title: Skeletal overgrowth in a pre-pubescent child treated with pan-FGFR inhibitor

    Journal: Heliyon

    doi: 10.1016/j.heliyon.2024.e30887

    A: Immunoblot analysis of MAPK and PI3K/AKT signaling in the presence of increasing doses of erdafitinib, growth factors- In untreated fibroblasts, ERK1/2 and AKT (both pAKT-T308 and pAKT-S473) are activated (in NC, normal control (no DMSO), 0 nM erdafitinib but with DMSO), Erdafitinib treatment at 100 nM dramatically reduce activated pERK1/2 and pAKT with no further reductions detected when fibroblasts were treated with 200 nM or 300 nM erdafitinib (3A-a) . Erdafitinib-inhibited pERK1/2 was not re-activated by the co-addition of IGF-I ( 3A-b ), FGF2 ( 3A-c ) or IGF-2 ( 3A-d ). AKT signaling pathway (pAKT-T308 and pAKT-S473) were consistently activated by IGF-I and IGF-2, but not by FGF2 ( 3A ), at all concentrations of erdafitinib tested (100 nM–300 nM) (see -uncropped.) .B: immunoblot analysis of signaling response in the presence of erdafitinib and growth factors over time- Fibroblasts were co-treated with 200nM erdafitinib and growth factors (IGF-1, IGF-2, FGF-2) for 0.5, 2, 24, 48 hours. Only with IGF-I treatment, activation of AKT was sustained for 48 hours. our data suggest, in our patient fibroblasts, that IGF-I activation of AKT signaling is sustained and independent of the FGFR signaling inhibitory effects of erdafitinib. (see -uncropped.).
    Figure Legend Snippet: A: Immunoblot analysis of MAPK and PI3K/AKT signaling in the presence of increasing doses of erdafitinib, growth factors- In untreated fibroblasts, ERK1/2 and AKT (both pAKT-T308 and pAKT-S473) are activated (in NC, normal control (no DMSO), 0 nM erdafitinib but with DMSO), Erdafitinib treatment at 100 nM dramatically reduce activated pERK1/2 and pAKT with no further reductions detected when fibroblasts were treated with 200 nM or 300 nM erdafitinib (3A-a) . Erdafitinib-inhibited pERK1/2 was not re-activated by the co-addition of IGF-I ( 3A-b ), FGF2 ( 3A-c ) or IGF-2 ( 3A-d ). AKT signaling pathway (pAKT-T308 and pAKT-S473) were consistently activated by IGF-I and IGF-2, but not by FGF2 ( 3A ), at all concentrations of erdafitinib tested (100 nM–300 nM) (see -uncropped.) .B: immunoblot analysis of signaling response in the presence of erdafitinib and growth factors over time- Fibroblasts were co-treated with 200nM erdafitinib and growth factors (IGF-1, IGF-2, FGF-2) for 0.5, 2, 24, 48 hours. Only with IGF-I treatment, activation of AKT was sustained for 48 hours. our data suggest, in our patient fibroblasts, that IGF-I activation of AKT signaling is sustained and independent of the FGFR signaling inhibitory effects of erdafitinib. (see -uncropped.).

    Techniques Used: Western Blot, Activation Assay

    rabbit monoclonal igg human phospho p44 42 mapk  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit monoclonal igg human phospho p44 42 mapk

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    1) Product Images from "Comprehensive proteogenomic characterization of rare kidney tumors"

    Article Title: Comprehensive proteogenomic characterization of rare kidney tumors

    Journal: Cell Reports Medicine

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    rabbit monoclonal igg human phospho p44 42 mapk  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 rabbit mab
    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Phospho P38 Mapk Thr180 Tyr182 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho p44 42 mapk erk1 2 thr202 tyr204 xp rabbit mab
    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Phospho P44 42 Mapk Erk1 2 Thr202 Tyr204 Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Rabbit Monoclonal Anti Phospho P38 Mapk Thr180 Tyr182 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a , b Volcano plot of microRNA analyzes of human samples, separated by sex for mature ( a ) and hairpin ( b ) microRNAs. We used DESEq2 for DE analysis and Benjamini-Hochberg for multiple test correction. The x -axis shows the log2 fold change in ALS vs CTR, whereas the y -axis shows the -log10 p-value. Orange and blue dots represent DEGs in females and males, respectively. miRNA-mediated regulation in ALS reveals a notable sex-dependent pattern. Male ALS patients exhibit a more pronounced downregulation of mature and hairpin miRNAs compared to females. c Heatmap showing mature miRNA expression changes for the identified ALS clusters. miRNA candidates significant in at least one condition were considered (p < 0.05). The scale (left-hand side) shows the range of log2 fold change values. d Volcano plot of differentially expressed proteins (DEP) in human samples (calculated with limma, two-sided test; multiple test correction with Benjamini-Hochberg). The x -axis shows the log2 fold change ALS and CTR, whereas the y -axis shows the -log10 p-values. Orange and blue dots represent DEGs in females and males, respectively. 379 DEPs in males and 251 in females. ANXA2 emerges as the sole protein downregulated in both sexes (p < 0.1). e , f Functional enrichment and unsupervised clustering using REVIGO, a tool used for summarizing Gene Ontology (GO) terms. Important pathway clusters are revealed in both sexes, including synaptic function, immune response, and ECM/cytoskeleton. Females ( e ) exhibited enrichment in transmembrane transport, lipid metabolism, development, catalytic activity, <t>and</t> <t>ERK1/2</t> signaling, while males ( f ) showed enrichment in cell metabolism and tyrosine kinase-related pathways. The size of the circles represents the number of genes in the GO-BP terms, and the color of each node represents the enrichment FDR values. g , h Uniform manifold approximation and projection (UMAP) of MOFA factor analysis for sex ( g ) and condition (ALS vs. CTR) ( h ). Sex ( g ) emerges as a robust differentiating factor when integrating transcriptomic, small RNA, and proteomic data. i MOFA correlation analysis displays which factors are dominated by which omic layer. Components of the representative factors 3 (males) and 12 (females) are displayed in feature-weight plots (bottom part). These factors are the ones that better correlate with disease conditions for each sex.
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    A: Immunoblot analysis of MAPK and PI3K/AKT signaling in the presence of increasing doses of erdafitinib, growth factors- In untreated <t>fibroblasts,</t> <t>ERK1/2</t> and AKT (both pAKT-T308 and pAKT-S473) are activated (in NC, normal control (no DMSO), 0 nM erdafitinib but with DMSO), Erdafitinib treatment at 100 nM dramatically reduce activated pERK1/2 and pAKT with no further reductions detected when fibroblasts were treated with 200 nM or 300 nM erdafitinib (3A-a) . Erdafitinib-inhibited pERK1/2 was not re-activated by the co-addition of IGF-I ( 3A-b ), FGF2 ( 3A-c ) or IGF-2 ( 3A-d ). AKT signaling pathway (pAKT-T308 and pAKT-S473) were consistently activated by IGF-I and IGF-2, but not by FGF2 ( 3A ), at all concentrations of erdafitinib tested (100 nM–300 nM) (see -uncropped.) .B: immunoblot analysis of signaling response in the presence of erdafitinib and growth factors over time- Fibroblasts were co-treated with 200nM erdafitinib and growth factors (IGF-1, IGF-2, FGF-2) for 0.5, 2, 24, 48 hours. Only with IGF-I treatment, activation of AKT was sustained for 48 hours. our data suggest, in our patient fibroblasts, that IGF-I activation of AKT signaling is sustained and independent of the FGFR signaling inhibitory effects of erdafitinib. (see -uncropped.).
    Anti Phospho P44 42 Mapk Erk1 2 Thr202 Tyr204 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: p38 MAPK inhibitor.

    Journal: Neural Regeneration Research

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    doi: 10.4103/1673-5374.391193

    Figure Lengend Snippet: Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: p38 MAPK inhibitor.

    Article Snippet: The following primary antibodies were used for staining: p38 MAPK rabbit polyclonal antibody (1:200, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), FTL rabbit polyclonal antibody (1:200, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673) and SAT1 rabbit polyclonal antibody (1:200, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK Thr180/Tyr182 rabbit mAb (1:200, Cell Signaling Technology, Cat# 4511); rabbit recombinant anti-GPx4 antibody (1:200, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 rabbit polyclonal antibody (1:200, Thermo Fisher Scientific, Cat# PA1-16893).

    Techniques: In Vivo, In Vitro

    p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced cell death. (A) The effect of different concentrations of glutamate on R28 cell viability over 24 hours ( n = 5). (B) Light microscopy was used to examine the effect of SB202190 on 10 mM glutamate-treated R28 cells. (C) Effect of SB202190 on the cell viability of R28 cells treated with 10 mM glutamate for 24 hours ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . control group (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. (D) The effect of SB202190 on the morphology of R28 cells was observed under a light microscope. Arrows indicate dead cells. Scale bars: 20 µm.

    Journal: Neural Regeneration Research

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    doi: 10.4103/1673-5374.391193

    Figure Lengend Snippet: p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced cell death. (A) The effect of different concentrations of glutamate on R28 cell viability over 24 hours ( n = 5). (B) Light microscopy was used to examine the effect of SB202190 on 10 mM glutamate-treated R28 cells. (C) Effect of SB202190 on the cell viability of R28 cells treated with 10 mM glutamate for 24 hours ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . control group (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. (D) The effect of SB202190 on the morphology of R28 cells was observed under a light microscope. Arrows indicate dead cells. Scale bars: 20 µm.

    Article Snippet: The following primary antibodies were used for staining: p38 MAPK rabbit polyclonal antibody (1:200, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), FTL rabbit polyclonal antibody (1:200, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673) and SAT1 rabbit polyclonal antibody (1:200, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK Thr180/Tyr182 rabbit mAb (1:200, Cell Signaling Technology, Cat# 4511); rabbit recombinant anti-GPx4 antibody (1:200, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 rabbit polyclonal antibody (1:200, Thermo Fisher Scientific, Cat# PA1-16893).

    Techniques: Light Microscopy, Comparison

    p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced ferroptosis. (A) Propidium iodide (PI; red) and Hoechst 33342 (blue) staining of R28 cells treated with glutamate, SB202190 and ferrostatin-1 as indicated for 24 hours. Scale bar: 100 µm. (B) Quantification of the results shown in A ( n = 5). (C) Transmission electron microscope images of R28 cells treated with glutamate and SB202190. Scale bar: 1 µm. (D) R28 cells treated as indicated were stained with the fluorescent probe Mito -FerroOrange to determine ferrous ion levels. FerroOrange undergoes an irreversible reaction with intracellular Fe 2+ in live cells, resulting in orange fluorescence emission. Ex: 543 nm and Em: 580 nm. Scale bar: 50 µm. (E) Quantification of mean density of FerroOrange fluorescence in the groups ( n = 5). (F) R28 cells treated as indicated were stained with Liperfluo for measurement of lipid peroxides. Liperfluo is selectively oxidized by lipid peroxides, and the oxidized form of Liperfluo exhibits strong green fluorescence at the cell membrane. Scale bar: 50 µm. (G) Comparison of mean density of Liperfluor fluorescence in the groups ( n = 5). (H, I) Malondialdehyde (MDA; H) and glutathione (GSH; I) levels in R28 cells after glutamate and SB202190 treatment ( n = 3). ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.

    Journal: Neural Regeneration Research

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    doi: 10.4103/1673-5374.391193

    Figure Lengend Snippet: p38 MAPK inhibitor SB202190 protects R28 cells from glutamate-induced ferroptosis. (A) Propidium iodide (PI; red) and Hoechst 33342 (blue) staining of R28 cells treated with glutamate, SB202190 and ferrostatin-1 as indicated for 24 hours. Scale bar: 100 µm. (B) Quantification of the results shown in A ( n = 5). (C) Transmission electron microscope images of R28 cells treated with glutamate and SB202190. Scale bar: 1 µm. (D) R28 cells treated as indicated were stained with the fluorescent probe Mito -FerroOrange to determine ferrous ion levels. FerroOrange undergoes an irreversible reaction with intracellular Fe 2+ in live cells, resulting in orange fluorescence emission. Ex: 543 nm and Em: 580 nm. Scale bar: 50 µm. (E) Quantification of mean density of FerroOrange fluorescence in the groups ( n = 5). (F) R28 cells treated as indicated were stained with Liperfluo for measurement of lipid peroxides. Liperfluo is selectively oxidized by lipid peroxides, and the oxidized form of Liperfluo exhibits strong green fluorescence at the cell membrane. Scale bar: 50 µm. (G) Comparison of mean density of Liperfluor fluorescence in the groups ( n = 5). (H, I) Malondialdehyde (MDA; H) and glutathione (GSH; I) levels in R28 cells after glutamate and SB202190 treatment ( n = 3). ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.

    Article Snippet: The following primary antibodies were used for staining: p38 MAPK rabbit polyclonal antibody (1:200, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), FTL rabbit polyclonal antibody (1:200, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673) and SAT1 rabbit polyclonal antibody (1:200, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK Thr180/Tyr182 rabbit mAb (1:200, Cell Signaling Technology, Cat# 4511); rabbit recombinant anti-GPx4 antibody (1:200, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 rabbit polyclonal antibody (1:200, Thermo Fisher Scientific, Cat# PA1-16893).

    Techniques: Staining, Transmission Assay, Microscopy, Fluorescence, Membrane, Comparison

    p38 MAPK inhibitor SB202190 regulates ferroptosis in a glutamate R28 cell model by regulating FTL and SAT1. (A) p38 MAPK, p-p38 MAPK, FTL and SAT1 protein expression in R28 cells treated with glutamate and SB202190 for 24 hours ( n = 3). (B–D) Quantification analysis of p-p38 MAPK/ p38 MAPK, FTL and SAT1 protein expression in R28 cells (n = 3). (E–I) Three days after intravitreal injection of NMDA and SB202190, paraffin sections were collected and processed for immunofluorescence experiments to measure the fluorescence intensity of p-p38 MAPK (E, F), FTL (G, H), and SAT1 (I, J) of the retinal sections in each group under a fluorescence microscope ( n = 3). Scale bars: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were repeated. FTL: Ferritin light chain; MAPK: mitogen activated protein kinases.

    Journal: Neural Regeneration Research

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    doi: 10.4103/1673-5374.391193

    Figure Lengend Snippet: p38 MAPK inhibitor SB202190 regulates ferroptosis in a glutamate R28 cell model by regulating FTL and SAT1. (A) p38 MAPK, p-p38 MAPK, FTL and SAT1 protein expression in R28 cells treated with glutamate and SB202190 for 24 hours ( n = 3). (B–D) Quantification analysis of p-p38 MAPK/ p38 MAPK, FTL and SAT1 protein expression in R28 cells (n = 3). (E–I) Three days after intravitreal injection of NMDA and SB202190, paraffin sections were collected and processed for immunofluorescence experiments to measure the fluorescence intensity of p-p38 MAPK (E, F), FTL (G, H), and SAT1 (I, J) of the retinal sections in each group under a fluorescence microscope ( n = 3). Scale bars: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were repeated. FTL: Ferritin light chain; MAPK: mitogen activated protein kinases.

    Article Snippet: The following primary antibodies were used for staining: p38 MAPK rabbit polyclonal antibody (1:200, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), FTL rabbit polyclonal antibody (1:200, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673) and SAT1 rabbit polyclonal antibody (1:200, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK Thr180/Tyr182 rabbit mAb (1:200, Cell Signaling Technology, Cat# 4511); rabbit recombinant anti-GPx4 antibody (1:200, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 rabbit polyclonal antibody (1:200, Thermo Fisher Scientific, Cat# PA1-16893).

    Techniques: Expressing, Injection, Immunofluorescence, Fluorescence, Microscopy, Comparison

    p38 MAPK inhibitor SB202190 alleviates lipid peroxidation by regulating the SLC7A11/GSH/GPx4 pathway. (A, B) Flow cytometry was used to detect the effect of SB202190 on lipid reactive oxygen species production in R28 cells treated as indicated for 24 hours ( n = 4). (C–E) SLC7A11 and GPx4 protein expressions in R28 cells were detected by western blotting ( n = 3). (F–I) Fluorescence microscopy was performed to evaluate the fluorescence intensity changes of and GPx4 (F, G) and SLC7A11 (H, I) in rat retinal sections after intraluminal injection of NMDA and SB202190 for 3 days ( n = 3). Scale bars in F and H: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. GPx4: Glutathione peroxidase 4; GSH: glutathione; MAPK: mitogen-activated protein kinase.

    Journal: Neural Regeneration Research

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    doi: 10.4103/1673-5374.391193

    Figure Lengend Snippet: p38 MAPK inhibitor SB202190 alleviates lipid peroxidation by regulating the SLC7A11/GSH/GPx4 pathway. (A, B) Flow cytometry was used to detect the effect of SB202190 on lipid reactive oxygen species production in R28 cells treated as indicated for 24 hours ( n = 4). (C–E) SLC7A11 and GPx4 protein expressions in R28 cells were detected by western blotting ( n = 3). (F–I) Fluorescence microscopy was performed to evaluate the fluorescence intensity changes of and GPx4 (F, G) and SLC7A11 (H, I) in rat retinal sections after intraluminal injection of NMDA and SB202190 for 3 days ( n = 3). Scale bars in F and H: 10 µm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. GPx4: Glutathione peroxidase 4; GSH: glutathione; MAPK: mitogen-activated protein kinase.

    Article Snippet: The following primary antibodies were used for staining: p38 MAPK rabbit polyclonal antibody (1:200, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), FTL rabbit polyclonal antibody (1:200, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673) and SAT1 rabbit polyclonal antibody (1:200, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK Thr180/Tyr182 rabbit mAb (1:200, Cell Signaling Technology, Cat# 4511); rabbit recombinant anti-GPx4 antibody (1:200, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 rabbit polyclonal antibody (1:200, Thermo Fisher Scientific, Cat# PA1-16893).

    Techniques: Flow Cytometry, Western Blot, Fluorescence, Microscopy, Injection, Comparison

    p38 MAPK inhibitor SB202190 attenuates N-methyl-D-aspartate (NMDA)-induced damage in the rat retina. (A, B) Qualitative observation (A) and quantitative analysis (B) of the effects of SB202190 on NMDA-induced retinal ganglion cell (RGC) injury in rat retina ( n = 5). Three days after intravitreal injection of NMDA and SB202190, retinal plating was performed. RGCs were fluorescently labeled with Brn3a antibody and surviving RGC exhibited strong green fluorescence under a fluorescence microscope. Scale bar: 50 μm. (C) Effects of SB202190 on retinal morphology in NMDA model rats. Hematoxylin and eosin staining was performed 3 days after intravitreal injection of NMDA and SB202190. Scale bar: 50 µm. (D) Effect of SB202190 on RGC density (cells/100 µm) in NMDA model rats 3 days after intravitreal injection ( n = 4). (E, F) Thickness of the retinal ganglion cell body complex (GCC) at 500, 1000, 1500, and 2000 µm from the optic disc 3 days after intravitreal injection of NMDA and SB202190. (G–I) Flash visual evoked potentials of rats 3 days after intravitreal injection of NMDA and SB202190 ( n = 4). GCL: Ganglion cell layer; INL: inner nuclear layer; IPL: inner plexiform layer; ONL: outer nuclear layer; RGC: retinal ganglion cell. Inner retina consists of GCL and IPL. ** P < 0.01, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.

    Journal: Neural Regeneration Research

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    doi: 10.4103/1673-5374.391193

    Figure Lengend Snippet: p38 MAPK inhibitor SB202190 attenuates N-methyl-D-aspartate (NMDA)-induced damage in the rat retina. (A, B) Qualitative observation (A) and quantitative analysis (B) of the effects of SB202190 on NMDA-induced retinal ganglion cell (RGC) injury in rat retina ( n = 5). Three days after intravitreal injection of NMDA and SB202190, retinal plating was performed. RGCs were fluorescently labeled with Brn3a antibody and surviving RGC exhibited strong green fluorescence under a fluorescence microscope. Scale bar: 50 μm. (C) Effects of SB202190 on retinal morphology in NMDA model rats. Hematoxylin and eosin staining was performed 3 days after intravitreal injection of NMDA and SB202190. Scale bar: 50 µm. (D) Effect of SB202190 on RGC density (cells/100 µm) in NMDA model rats 3 days after intravitreal injection ( n = 4). (E, F) Thickness of the retinal ganglion cell body complex (GCC) at 500, 1000, 1500, and 2000 µm from the optic disc 3 days after intravitreal injection of NMDA and SB202190. (G–I) Flash visual evoked potentials of rats 3 days after intravitreal injection of NMDA and SB202190 ( n = 4). GCL: Ganglion cell layer; INL: inner nuclear layer; IPL: inner plexiform layer; ONL: outer nuclear layer; RGC: retinal ganglion cell. Inner retina consists of GCL and IPL. ** P < 0.01, **** P < 0.0001 (one-way analysis of variance followed by Tukey’s multiple comparison test). Data are expressed as the means ± SD. At least three independent experiments were performed. MAPK: Mitogen-activated protein kinase.

    Article Snippet: The following primary antibodies were used for staining: p38 MAPK rabbit polyclonal antibody (1:200, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), FTL rabbit polyclonal antibody (1:200, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673) and SAT1 rabbit polyclonal antibody (1:200, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK Thr180/Tyr182 rabbit mAb (1:200, Cell Signaling Technology, Cat# 4511); rabbit recombinant anti-GPx4 antibody (1:200, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 rabbit polyclonal antibody (1:200, Thermo Fisher Scientific, Cat# PA1-16893).

    Techniques: Injection, Labeling, Fluorescence, Microscopy, Staining, Comparison

    A schematic model for the mechanism of p38 MAPK inhibitor SB202190 in alleviating ferroptosis in the glutamate-induced retinal excitotoxic glaucoma model. The p38 MAPK inhibitor sB202190 inhibits ferroptosis in the glutamatergic excitotoxicity glaucoma model by upregulation of ferritin light chain (FTL), downregulation of SAT1 and regulation of the SLC7A11/GSH/GPx4 signaling pathway. GPx4: Glutathione peroxidase 4; GSH: glutathione; GSSG: glutathione disulfide; MAPK: mitogen-activated protein kinase; ROS: reactive oxygen species.

    Journal: Neural Regeneration Research

    Article Title: p38 MAPK inhibitor SB202190 suppresses ferroptosis in the glutamate-induced retinal excitotoxicity glaucoma model

    doi: 10.4103/1673-5374.391193

    Figure Lengend Snippet: A schematic model for the mechanism of p38 MAPK inhibitor SB202190 in alleviating ferroptosis in the glutamate-induced retinal excitotoxic glaucoma model. The p38 MAPK inhibitor sB202190 inhibits ferroptosis in the glutamatergic excitotoxicity glaucoma model by upregulation of ferritin light chain (FTL), downregulation of SAT1 and regulation of the SLC7A11/GSH/GPx4 signaling pathway. GPx4: Glutathione peroxidase 4; GSH: glutathione; GSSG: glutathione disulfide; MAPK: mitogen-activated protein kinase; ROS: reactive oxygen species.

    Article Snippet: The following primary antibodies were used for staining: p38 MAPK rabbit polyclonal antibody (1:200, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), FTL rabbit polyclonal antibody (1:200, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673) and SAT1 rabbit polyclonal antibody (1:200, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK Thr180/Tyr182 rabbit mAb (1:200, Cell Signaling Technology, Cat# 4511); rabbit recombinant anti-GPx4 antibody (1:200, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 rabbit polyclonal antibody (1:200, Thermo Fisher Scientific, Cat# PA1-16893).

    Techniques:

    a , b Volcano plot of microRNA analyzes of human samples, separated by sex for mature ( a ) and hairpin ( b ) microRNAs. We used DESEq2 for DE analysis and Benjamini-Hochberg for multiple test correction. The x -axis shows the log2 fold change in ALS vs CTR, whereas the y -axis shows the -log10 p-value. Orange and blue dots represent DEGs in females and males, respectively. miRNA-mediated regulation in ALS reveals a notable sex-dependent pattern. Male ALS patients exhibit a more pronounced downregulation of mature and hairpin miRNAs compared to females. c Heatmap showing mature miRNA expression changes for the identified ALS clusters. miRNA candidates significant in at least one condition were considered (p < 0.05). The scale (left-hand side) shows the range of log2 fold change values. d Volcano plot of differentially expressed proteins (DEP) in human samples (calculated with limma, two-sided test; multiple test correction with Benjamini-Hochberg). The x -axis shows the log2 fold change ALS and CTR, whereas the y -axis shows the -log10 p-values. Orange and blue dots represent DEGs in females and males, respectively. 379 DEPs in males and 251 in females. ANXA2 emerges as the sole protein downregulated in both sexes (p < 0.1). e , f Functional enrichment and unsupervised clustering using REVIGO, a tool used for summarizing Gene Ontology (GO) terms. Important pathway clusters are revealed in both sexes, including synaptic function, immune response, and ECM/cytoskeleton. Females ( e ) exhibited enrichment in transmembrane transport, lipid metabolism, development, catalytic activity, and ERK1/2 signaling, while males ( f ) showed enrichment in cell metabolism and tyrosine kinase-related pathways. The size of the circles represents the number of genes in the GO-BP terms, and the color of each node represents the enrichment FDR values. g , h Uniform manifold approximation and projection (UMAP) of MOFA factor analysis for sex ( g ) and condition (ALS vs. CTR) ( h ). Sex ( g ) emerges as a robust differentiating factor when integrating transcriptomic, small RNA, and proteomic data. i MOFA correlation analysis displays which factors are dominated by which omic layer. Components of the representative factors 3 (males) and 12 (females) are displayed in feature-weight plots (bottom part). These factors are the ones that better correlate with disease conditions for each sex.

    Journal: Nature Communications

    Article Title: Multiomic ALS signatures highlight subclusters and sex differences suggesting the MAPK pathway as therapeutic target

    doi: 10.1038/s41467-024-49196-y

    Figure Lengend Snippet: a , b Volcano plot of microRNA analyzes of human samples, separated by sex for mature ( a ) and hairpin ( b ) microRNAs. We used DESEq2 for DE analysis and Benjamini-Hochberg for multiple test correction. The x -axis shows the log2 fold change in ALS vs CTR, whereas the y -axis shows the -log10 p-value. Orange and blue dots represent DEGs in females and males, respectively. miRNA-mediated regulation in ALS reveals a notable sex-dependent pattern. Male ALS patients exhibit a more pronounced downregulation of mature and hairpin miRNAs compared to females. c Heatmap showing mature miRNA expression changes for the identified ALS clusters. miRNA candidates significant in at least one condition were considered (p < 0.05). The scale (left-hand side) shows the range of log2 fold change values. d Volcano plot of differentially expressed proteins (DEP) in human samples (calculated with limma, two-sided test; multiple test correction with Benjamini-Hochberg). The x -axis shows the log2 fold change ALS and CTR, whereas the y -axis shows the -log10 p-values. Orange and blue dots represent DEGs in females and males, respectively. 379 DEPs in males and 251 in females. ANXA2 emerges as the sole protein downregulated in both sexes (p < 0.1). e , f Functional enrichment and unsupervised clustering using REVIGO, a tool used for summarizing Gene Ontology (GO) terms. Important pathway clusters are revealed in both sexes, including synaptic function, immune response, and ECM/cytoskeleton. Females ( e ) exhibited enrichment in transmembrane transport, lipid metabolism, development, catalytic activity, and ERK1/2 signaling, while males ( f ) showed enrichment in cell metabolism and tyrosine kinase-related pathways. The size of the circles represents the number of genes in the GO-BP terms, and the color of each node represents the enrichment FDR values. g , h Uniform manifold approximation and projection (UMAP) of MOFA factor analysis for sex ( g ) and condition (ALS vs. CTR) ( h ). Sex ( g ) emerges as a robust differentiating factor when integrating transcriptomic, small RNA, and proteomic data. i MOFA correlation analysis displays which factors are dominated by which omic layer. Components of the representative factors 3 (males) and 12 (females) are displayed in feature-weight plots (bottom part). These factors are the ones that better correlate with disease conditions for each sex.

    Article Snippet: The antibodies utilized were rabbit monoclonal anti-pMEK2 (1:2000, Cell Signaling Technology; Danvers, MA, USA, RRID: AB_490903), rabbit monoclonal anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (1:2000, Cell Signaling Technology, Danvers, MA, USA,; RRID: AB_2315112), rabbit polyclonal anti-human SOD1 (1:1000, StressMarq Biosciences, Victoria, Canada; RRID: AB_2704217), rabbit polyclonal anti-ubiquitin (1:1000, Abcam, Cambridge, UK; RRID: AB_306069), and mouse monoclonal anti-SQSTM1/p62 (p62) (1:500, Abcam, Cambridge, United Kingdom, RRID: AB_945626).

    Techniques: Expressing, Functional Assay, Activity Assay

    Journal: iScience

    Article Title: Oxygen tension-dependent variability in the cancer cell kinome impacts signaling pathways and response to targeted therapies

    doi: 10.1016/j.isci.2024.110068

    Figure Lengend Snippet:

    Article Snippet: Rabbit monoclonal anti-phospho-p44/42 MAPK (T202/Y204) , Cell Signaling Technology , Cat# 9101; RRID:AB_331646.

    Techniques: Recombinant, Gentle, Lysis, Protease Inhibitor, BrdU Cell Proliferation Assay, Caspase-3 Assay, RNA Sequencing Assay, Mass Spectrometry, Transgenic Assay, TaqMan Assay, Software, Modification, Flow Cytometry, Real-time Polymerase Chain Reaction, Isolation

    A: Immunoblot analysis of MAPK and PI3K/AKT signaling in the presence of increasing doses of erdafitinib, growth factors- In untreated fibroblasts, ERK1/2 and AKT (both pAKT-T308 and pAKT-S473) are activated (in NC, normal control (no DMSO), 0 nM erdafitinib but with DMSO), Erdafitinib treatment at 100 nM dramatically reduce activated pERK1/2 and pAKT with no further reductions detected when fibroblasts were treated with 200 nM or 300 nM erdafitinib (3A-a) . Erdafitinib-inhibited pERK1/2 was not re-activated by the co-addition of IGF-I ( 3A-b ), FGF2 ( 3A-c ) or IGF-2 ( 3A-d ). AKT signaling pathway (pAKT-T308 and pAKT-S473) were consistently activated by IGF-I and IGF-2, but not by FGF2 ( 3A ), at all concentrations of erdafitinib tested (100 nM–300 nM) (see -uncropped.) .B: immunoblot analysis of signaling response in the presence of erdafitinib and growth factors over time- Fibroblasts were co-treated with 200nM erdafitinib and growth factors (IGF-1, IGF-2, FGF-2) for 0.5, 2, 24, 48 hours. Only with IGF-I treatment, activation of AKT was sustained for 48 hours. our data suggest, in our patient fibroblasts, that IGF-I activation of AKT signaling is sustained and independent of the FGFR signaling inhibitory effects of erdafitinib. (see -uncropped.).

    Journal: Heliyon

    Article Title: Skeletal overgrowth in a pre-pubescent child treated with pan-FGFR inhibitor

    doi: 10.1016/j.heliyon.2024.e30887

    Figure Lengend Snippet: A: Immunoblot analysis of MAPK and PI3K/AKT signaling in the presence of increasing doses of erdafitinib, growth factors- In untreated fibroblasts, ERK1/2 and AKT (both pAKT-T308 and pAKT-S473) are activated (in NC, normal control (no DMSO), 0 nM erdafitinib but with DMSO), Erdafitinib treatment at 100 nM dramatically reduce activated pERK1/2 and pAKT with no further reductions detected when fibroblasts were treated with 200 nM or 300 nM erdafitinib (3A-a) . Erdafitinib-inhibited pERK1/2 was not re-activated by the co-addition of IGF-I ( 3A-b ), FGF2 ( 3A-c ) or IGF-2 ( 3A-d ). AKT signaling pathway (pAKT-T308 and pAKT-S473) were consistently activated by IGF-I and IGF-2, but not by FGF2 ( 3A ), at all concentrations of erdafitinib tested (100 nM–300 nM) (see -uncropped.) .B: immunoblot analysis of signaling response in the presence of erdafitinib and growth factors over time- Fibroblasts were co-treated with 200nM erdafitinib and growth factors (IGF-1, IGF-2, FGF-2) for 0.5, 2, 24, 48 hours. Only with IGF-I treatment, activation of AKT was sustained for 48 hours. our data suggest, in our patient fibroblasts, that IGF-I activation of AKT signaling is sustained and independent of the FGFR signaling inhibitory effects of erdafitinib. (see -uncropped.).

    Article Snippet: Antibodies used were anti-phospho-AKT(Ser 473), anti-phospho-AKT(Thr308), and anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) rabbit mAb (Cell Signaling Technology, Danvers, MA), anti-Akt (pan) Rabbit mAb, p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb(Cell Signaling Technology, Danvers, MA), anti-PARP rabbit mAb and Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb (Cell Signaling Technology, Danvers, MA), anti-alpha tubulin monoclonal horseradish peroxidase conjugate (Cell Signaling Technology, Danvers, MA), Secondary antibodies (horseradish peroxidase-linked anti-rabbit IgG and anti-mouse IgG antibodies) were obtained from Amersham Biosciences (Uppsala, Sweden).

    Techniques: Western Blot, Activation Assay

    Journal: Cell Reports Medicine

    Article Title: Comprehensive proteogenomic characterization of rare kidney tumors

    doi: 10.1016/j.xcrm.2024.101547

    Figure Lengend Snippet:

    Article Snippet: Rabbit Monoclonal IgG Human Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody , Cell Signaling Technology , Catalog: 4376.

    Techniques: Software