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
    https://www.bioz.com/result/phospho p38 mapk thr180 tyr182 rabbit mab/product/Cell Signaling Technology Inc
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    phospho p38 mapk thr180 tyr182 rabbit mab - by Bioz Stars, 2024-09
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    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 p38 mapk thr180 tyr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182
    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, <t>P38,</t> p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
    Phospho P38 Mapk Thr180 Tyr182, 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 "Human neural stem cell-derived artificial organelles to improve oxidative phosphorylation"

    Article Title: Human neural stem cell-derived artificial organelles to improve oxidative phosphorylation

    Journal: Nature Communications

    doi: 10.1038/s41467-024-52171-2

    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, P38, p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
    Figure Legend Snippet: a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, P38, p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.

    Techniques Used: RNA Sequencing Assay, TUNEL Assay, Lactate Dehydrogenase Assay, Western Blot, Control

    p38 mapk phospho thr180 tyr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p38 mapk phospho thr180 tyr182
    P38 Mapk Phospho Thr180 Tyr182, 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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    phospho p38 mapk antibody thr180 tyr182 d3f9  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk antibody thr180 tyr182 d3f9
    Phospho P38 Mapk Antibody Thr180 Tyr182 D3f9, 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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    phospho p38 mapk thr180 tyr182 polyclonal antibody  (Proteintech)

     
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    Proteintech phospho p38 mapk thr180 tyr182 polyclonal antibody
    Phospho P38 Mapk Thr180 Tyr182 Polyclonal Antibody, supplied by Proteintech, 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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    phospho p38 mapk thr180 tyr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182
    Phospho P38 Mapk Thr180 Tyr182, 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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    phospho p38 mapk thr180 tyr182 primary antibodies  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 primary antibodies
    Phospho P38 Mapk Thr180 Tyr182 Primary Antibodies, 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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    phospho p38 mapk thr180 tyr182 d3f9 xp rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 d3f9 xp rabbit mab
    Phospho P38 Mapk Thr180 Tyr182 D3f9 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
    https://www.bioz.com/result/phospho p38 mapk thr180 tyr182 d3f9 xp rabbit mab/product/Cell Signaling Technology Inc
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    phospho p38 mapk thr180 tyr182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182
    Activation of TGFβ signaling enhances p63 recruitment to DNA via <t>ALK5/p38</t> kinase-dependent phosphorylation. (A-B) Sphere formation assay of HCC1954 cells in the presence or absence of TGFβ as indicated. Cells were transfected with non-targeting control (sictrl) siRNA or with siRNAs specific against all p63 isoforms (A) or specific against the ΔNp63 isoforms (B) . Cells were cultured in stem cell medium in 96-well ultra-low attachment plates. Sphere numbers per well were counted under microscopy. (C) ChIP-qPCR showing the effect of ALK5 kinase or MEK1/2 kinase inhibition on the TGFβ-induced recruitment of p63 to DNA. Values are expressed as relative fold-change corresponding to the enrichment of p63 antibody in each gene locus under treatment conditions divided by the enrichment in the control condition (ctrl). (D-E) TGFβ stimulation induced ALK5- and p38-dependent phosphorylation of ΔNp63 at Ser66/Ser68. Immunoblotting (IB) analysis of MCF10A MII cells, treated with the indicated kinase inhibitors or DMSO (ctrl) in the presence of TGFβ for 6 h. (F) Cycloheximide (CHX) chase experiment in MCF10A MII cells. Lysates of cells treated or not with p38 inhibitor (SB203580) or DMSO (ctrl) in the presence or not of TGFβ for 6 h were analyzed by IB with the indicated antibodies. In panels D-F, one of four independent experiments with similar results, is shown. The intensities of the p-p63 Ser66/68, p63 and tubulin bands from each of the four independent experiments were quantified and the values were used to calculate the ratio of p-p63/p63, presented between the corresponding immunoblots. (G) ChIP-qPCR showing the effect of p38 inhibition on the TGFβ-induced recruitment of p63 to DNA. Graphs presented in panels A-C and G show results of three independent experiments as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001. The dots in the graphs of panels C and G represent the individual values from each of the three independent ChIP experiments
    Phospho P38 Mapk Thr180 Tyr182, 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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    Images

    1) Product Images from "ΔNp63 bookmarks and creates an accessible epigenetic environment for TGFβ-induced cancer cell stemness and invasiveness"

    Article Title: ΔNp63 bookmarks and creates an accessible epigenetic environment for TGFβ-induced cancer cell stemness and invasiveness

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-024-01794-5

    Activation of TGFβ signaling enhances p63 recruitment to DNA via ALK5/p38 kinase-dependent phosphorylation. (A-B) Sphere formation assay of HCC1954 cells in the presence or absence of TGFβ as indicated. Cells were transfected with non-targeting control (sictrl) siRNA or with siRNAs specific against all p63 isoforms (A) or specific against the ΔNp63 isoforms (B) . Cells were cultured in stem cell medium in 96-well ultra-low attachment plates. Sphere numbers per well were counted under microscopy. (C) ChIP-qPCR showing the effect of ALK5 kinase or MEK1/2 kinase inhibition on the TGFβ-induced recruitment of p63 to DNA. Values are expressed as relative fold-change corresponding to the enrichment of p63 antibody in each gene locus under treatment conditions divided by the enrichment in the control condition (ctrl). (D-E) TGFβ stimulation induced ALK5- and p38-dependent phosphorylation of ΔNp63 at Ser66/Ser68. Immunoblotting (IB) analysis of MCF10A MII cells, treated with the indicated kinase inhibitors or DMSO (ctrl) in the presence of TGFβ for 6 h. (F) Cycloheximide (CHX) chase experiment in MCF10A MII cells. Lysates of cells treated or not with p38 inhibitor (SB203580) or DMSO (ctrl) in the presence or not of TGFβ for 6 h were analyzed by IB with the indicated antibodies. In panels D-F, one of four independent experiments with similar results, is shown. The intensities of the p-p63 Ser66/68, p63 and tubulin bands from each of the four independent experiments were quantified and the values were used to calculate the ratio of p-p63/p63, presented between the corresponding immunoblots. (G) ChIP-qPCR showing the effect of p38 inhibition on the TGFβ-induced recruitment of p63 to DNA. Graphs presented in panels A-C and G show results of three independent experiments as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001. The dots in the graphs of panels C and G represent the individual values from each of the three independent ChIP experiments
    Figure Legend Snippet: Activation of TGFβ signaling enhances p63 recruitment to DNA via ALK5/p38 kinase-dependent phosphorylation. (A-B) Sphere formation assay of HCC1954 cells in the presence or absence of TGFβ as indicated. Cells were transfected with non-targeting control (sictrl) siRNA or with siRNAs specific against all p63 isoforms (A) or specific against the ΔNp63 isoforms (B) . Cells were cultured in stem cell medium in 96-well ultra-low attachment plates. Sphere numbers per well were counted under microscopy. (C) ChIP-qPCR showing the effect of ALK5 kinase or MEK1/2 kinase inhibition on the TGFβ-induced recruitment of p63 to DNA. Values are expressed as relative fold-change corresponding to the enrichment of p63 antibody in each gene locus under treatment conditions divided by the enrichment in the control condition (ctrl). (D-E) TGFβ stimulation induced ALK5- and p38-dependent phosphorylation of ΔNp63 at Ser66/Ser68. Immunoblotting (IB) analysis of MCF10A MII cells, treated with the indicated kinase inhibitors or DMSO (ctrl) in the presence of TGFβ for 6 h. (F) Cycloheximide (CHX) chase experiment in MCF10A MII cells. Lysates of cells treated or not with p38 inhibitor (SB203580) or DMSO (ctrl) in the presence or not of TGFβ for 6 h were analyzed by IB with the indicated antibodies. In panels D-F, one of four independent experiments with similar results, is shown. The intensities of the p-p63 Ser66/68, p63 and tubulin bands from each of the four independent experiments were quantified and the values were used to calculate the ratio of p-p63/p63, presented between the corresponding immunoblots. (G) ChIP-qPCR showing the effect of p38 inhibition on the TGFβ-induced recruitment of p63 to DNA. Graphs presented in panels A-C and G show results of three independent experiments as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001. The dots in the graphs of panels C and G represent the individual values from each of the three independent ChIP experiments

    Techniques Used: Activation Assay, Tube Formation Assay, Transfection, Control, Cell Culture, Microscopy, Inhibition, Western Blot

    Activation of SMAD2 and SMAD3 transcription factors drives the p63 selectivity on histone modulation complexes. (A) Effect of ALK5 inhibition on p63/p300 interaction. Lysates of MCF10A MII cells treated with ALK5 kinase inhibitor (SB505124) or not (control) in the presence of TGFβ stimulation were subjected to IP with p300-specific antibody or IgG control, and analyzed by IB with the indicated antibodies. ( B ) Effect of p38 inhibition on p63/SMAD2/3 interaction. Lysates of MCF10A MII cells treated with p38 kinase inhibitor (SB203580) or not (control) in the presence of TGFβ stimulation were subjected to IP with SMAD2/3 antibody or IgG control, and analyzed by IB with the indicated antibodies. (C) Effect of SMAD2/3 depletion on p63/p300 interaction. MCF10A MII cells transfected with non-targeting control (sictrl) siRNA or with siRNA specific against SMAD2 and SMAD3 were incubated in starvation medium and treated or not with TGFβ for 6 h. Cell lysates were subjected to IP with p300-specific antibody or IgG control, and analyzed by IB with the indicated antibodies. (D) Effect of p63 depletion on SMAD3/p300 interaction. MCF10A MII cells transfected with non-targeting control (sictrl) siRNA or with siRNA specific against all p63 isoforms were incubated in starvation medium overnight and treated or not with TGFβ for 45 min. Cell lysates were subjected to IP with a p300-specific antibody or IgG control, and analyzed by IB with the indicated antibodies. (E , F) Effect of SMAD2/3 depletion on p300 recruitment to chromatin (E) and H3K27ac (F) . MCF10A MII cells transfected with siRNAs and starved as in panel C were treated or not with TGFβ for 24 h. Cell lysates were subjected to ChIP with p300 (E) or H3K27ac (F) antibodies and subsequent qPCR analysis. Graphs presented in panels E-F show results of three independent experiments as mean ± SD; * P < 0.05, ** P < 0.01. The dots represent the individual values from each of the three independent experiments. (G) Effect of SMAD2/3 depletion on p63/NCOR2 interaction. MCF10A MII cells transfected with siRNAs and starved as in panel A were treated or not with TGFβ for 6 h. Cell lysates were subjected to IP with p63 and analyzed by IB with the indicated antibodies. One of three independent experiments with similar results, is shown. ( H ) Graph illustrating the relative IP NCOR2/input as average values from the quantification of three independent experiments performed as described in panel G. The dots represent the individual values from each of the three independent experiments
    Figure Legend Snippet: Activation of SMAD2 and SMAD3 transcription factors drives the p63 selectivity on histone modulation complexes. (A) Effect of ALK5 inhibition on p63/p300 interaction. Lysates of MCF10A MII cells treated with ALK5 kinase inhibitor (SB505124) or not (control) in the presence of TGFβ stimulation were subjected to IP with p300-specific antibody or IgG control, and analyzed by IB with the indicated antibodies. ( B ) Effect of p38 inhibition on p63/SMAD2/3 interaction. Lysates of MCF10A MII cells treated with p38 kinase inhibitor (SB203580) or not (control) in the presence of TGFβ stimulation were subjected to IP with SMAD2/3 antibody or IgG control, and analyzed by IB with the indicated antibodies. (C) Effect of SMAD2/3 depletion on p63/p300 interaction. MCF10A MII cells transfected with non-targeting control (sictrl) siRNA or with siRNA specific against SMAD2 and SMAD3 were incubated in starvation medium and treated or not with TGFβ for 6 h. Cell lysates were subjected to IP with p300-specific antibody or IgG control, and analyzed by IB with the indicated antibodies. (D) Effect of p63 depletion on SMAD3/p300 interaction. MCF10A MII cells transfected with non-targeting control (sictrl) siRNA or with siRNA specific against all p63 isoforms were incubated in starvation medium overnight and treated or not with TGFβ for 45 min. Cell lysates were subjected to IP with a p300-specific antibody or IgG control, and analyzed by IB with the indicated antibodies. (E , F) Effect of SMAD2/3 depletion on p300 recruitment to chromatin (E) and H3K27ac (F) . MCF10A MII cells transfected with siRNAs and starved as in panel C were treated or not with TGFβ for 24 h. Cell lysates were subjected to ChIP with p300 (E) or H3K27ac (F) antibodies and subsequent qPCR analysis. Graphs presented in panels E-F show results of three independent experiments as mean ± SD; * P < 0.05, ** P < 0.01. The dots represent the individual values from each of the three independent experiments. (G) Effect of SMAD2/3 depletion on p63/NCOR2 interaction. MCF10A MII cells transfected with siRNAs and starved as in panel A were treated or not with TGFβ for 6 h. Cell lysates were subjected to IP with p63 and analyzed by IB with the indicated antibodies. One of three independent experiments with similar results, is shown. ( H ) Graph illustrating the relative IP NCOR2/input as average values from the quantification of three independent experiments performed as described in panel G. The dots represent the individual values from each of the three independent experiments

    Techniques Used: Activation Assay, Inhibition, Control, Transfection, Incubation

    Schematic illustration of the effect of active TGFβ signaling on ΔNp63-dependent transcription. (A) In the absence of active TGFβ pathway, ΔNp63 is bound to NURD/PRC2 and NCOR/SMRT/HDAC3 complexes on TGFβ/SMAD target regulatory genomic loci. These regions showing high H3K4me1 are bookmarked for transcription by ΔNp63; however, the presence of the H3K27 tri-methylation mark results in condensed chromatin and inactive transcription. (B) Activation of TGFβ signaling leads to phosphorylation of ΔNp63 at Ser66/68 via p38 MAPK, nuclear translocation of SMAD2/3 transcription factors and complex formation between ΔNp63, SMAD2/3 and p300. p300 catalyzes the acetylation of K27 on H3, which promotes chromatin accessibility and activation of gene transcription favoring cancer cell stemness and invasiveness. Dynamic interactions with chromatin are shown with two anti-parallel arrows. Arrow thickness correlates to the prevalent interaction (association or dissociation). Created with BioRender.com
    Figure Legend Snippet: Schematic illustration of the effect of active TGFβ signaling on ΔNp63-dependent transcription. (A) In the absence of active TGFβ pathway, ΔNp63 is bound to NURD/PRC2 and NCOR/SMRT/HDAC3 complexes on TGFβ/SMAD target regulatory genomic loci. These regions showing high H3K4me1 are bookmarked for transcription by ΔNp63; however, the presence of the H3K27 tri-methylation mark results in condensed chromatin and inactive transcription. (B) Activation of TGFβ signaling leads to phosphorylation of ΔNp63 at Ser66/68 via p38 MAPK, nuclear translocation of SMAD2/3 transcription factors and complex formation between ΔNp63, SMAD2/3 and p300. p300 catalyzes the acetylation of K27 on H3, which promotes chromatin accessibility and activation of gene transcription favoring cancer cell stemness and invasiveness. Dynamic interactions with chromatin are shown with two anti-parallel arrows. Arrow thickness correlates to the prevalent interaction (association or dissociation). Created with BioRender.com

    Techniques Used: Methylation, Activation Assay, Translocation Assay

    phospho p38 mapk thr180 tyr182  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182
    A Immunoblotting of caspases in A549 and H1650 with ICAM-1 knockdown and re-expression of WT ICAM-1, ICAM-1Δ8-22 or ICAM-1ΔCyto. B Tyrosine phosphorylation of ICAM-1 and co-immunoprecipitation of SHP-2 with ICAM-1 in the indicated cells. C Phosphorylation of Akt (Ser473), ERK1/2 (Thr202/Tyr204), JNK (Thr183/Tyr185) and <t>p38</t> <t>(Thr180/Tyr182)</t> in A549 and H1650 cells in the indicated cells. Total Akt, ERK1/2, JNK, p38 and β-actin are shown in the lower rows. A representative result of three independent experiments is shown.
    Phospho P38 Mapk Thr180 Tyr182, 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 "Neoplastic ICAM-1 protects lung carcinoma from apoptosis through ligation of fibrinogen"

    Article Title: Neoplastic ICAM-1 protects lung carcinoma from apoptosis through ligation of fibrinogen

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-024-06989-9

    A Immunoblotting of caspases in A549 and H1650 with ICAM-1 knockdown and re-expression of WT ICAM-1, ICAM-1Δ8-22 or ICAM-1ΔCyto. B Tyrosine phosphorylation of ICAM-1 and co-immunoprecipitation of SHP-2 with ICAM-1 in the indicated cells. C Phosphorylation of Akt (Ser473), ERK1/2 (Thr202/Tyr204), JNK (Thr183/Tyr185) and p38 (Thr180/Tyr182) in A549 and H1650 cells in the indicated cells. Total Akt, ERK1/2, JNK, p38 and β-actin are shown in the lower rows. A representative result of three independent experiments is shown.
    Figure Legend Snippet: A Immunoblotting of caspases in A549 and H1650 with ICAM-1 knockdown and re-expression of WT ICAM-1, ICAM-1Δ8-22 or ICAM-1ΔCyto. B Tyrosine phosphorylation of ICAM-1 and co-immunoprecipitation of SHP-2 with ICAM-1 in the indicated cells. C Phosphorylation of Akt (Ser473), ERK1/2 (Thr202/Tyr204), JNK (Thr183/Tyr185) and p38 (Thr180/Tyr182) in A549 and H1650 cells in the indicated cells. Total Akt, ERK1/2, JNK, p38 and β-actin are shown in the lower rows. A representative result of three independent experiments is shown.

    Techniques Used: Western Blot, Knockdown, Expressing, Immunoprecipitation

    A ICAM-1 is tyrosine phosphorylated upon ligation with FGG through motif Lys8-Ser22, followed by association with SHP-2, activation of ERK1/2 and Akt and restraint of JNK and p38 MAPK signaling, and consequently inhibits caspase-9/3 activation and prevents cell apoptosis. B Abolishing ICAM-1–FGG interaction disrupts the anti-apoptotic signaling and induces cancer cell death.
    Figure Legend Snippet: A ICAM-1 is tyrosine phosphorylated upon ligation with FGG through motif Lys8-Ser22, followed by association with SHP-2, activation of ERK1/2 and Akt and restraint of JNK and p38 MAPK signaling, and consequently inhibits caspase-9/3 activation and prevents cell apoptosis. B Abolishing ICAM-1–FGG interaction disrupts the anti-apoptotic signaling and induces cancer cell death.

    Techniques Used: Ligation, Activation Assay

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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.
    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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    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, <t>P38,</t> p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
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    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, <t>P38,</t> p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
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    phospho p38 mapk antibody thr180 tyr182 d3f9  (Cell Signaling Technology Inc)
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    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, <t>P38,</t> p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
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    phospho p38 mapk thr180 tyr182 polyclonal antibody  (Proteintech)
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    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, <t>P38,</t> p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
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    phospho p38 mapk thr180 tyr182 primary antibodies  (Cell Signaling Technology Inc)
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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 primary antibodies
    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, <t>P38,</t> p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
    Phospho P38 Mapk Thr180 Tyr182 Primary Antibodies, 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/phospho p38 mapk thr180 tyr182 primary antibodies/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 mapk thr180 tyr182 primary antibodies - by Bioz Stars, 2024-09
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    		  Buy from Supplier
    phospho p38 mapk thr180 tyr182 d3f9 xp rabbit mab  (Cell Signaling Technology Inc)
    86
    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 d3f9 xp rabbit mab
    a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, <t>P38,</t> p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.
    Phospho P38 Mapk Thr180 Tyr182 D3f9 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
    https://www.bioz.com/result/phospho p38 mapk thr180 tyr182 d3f9 xp rabbit mab/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 mapk thr180 tyr182 d3f9 xp rabbit mab - by Bioz Stars, 2024-09
    86/100 stars
    		  Buy from Supplier

    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 Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, P38, p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.

    Journal: Nature Communications

    Article Title: Human neural stem cell-derived artificial organelles to improve oxidative phosphorylation

    doi: 10.1038/s41467-024-52171-2

    Figure Lengend Snippet: a Dimensionality reduction plot for visualizing single-nucleus RNA sequencing (SNS) data using UMAP of neural cells after exposure to H 2 O 2 with or without SAOs. b Gene set enrichment analysis was performed in cluster 9 and other clusters. c Dimensionality reduction for visualizing SNS data using UMAP of neural cells. d Gene set enrichment analysis was performed in apoptosis cluster and other clusters. e The morphology of human mix neural cells. f Representative images and quantification of TUNEL+ cells in neural cells ( n = 12). g Cell viability was determined by lactate dehydrogenase (LDH) release assay ( n = 3). h Immunoblot of ERK, p-ERK, P38, p-P38, JNK, p-JNK in neural cells. GAPDH was used as a loading control. i Quantitative analysis of ERK, p-ERK, P38, p-P38, JNK, p-JNK ( n = 3). Transcriptomic analysis was performed using a sample size of three ( n = 3) for each group in ( a – d ). All experiments were conducted in triplicate unless otherwise indicated. Statistical analysis, one-way ANOVA ( f , g , i ). Error bars: mean ± s.e.m. Scale bar, 200 μm ( e ), 100 μm ( f ). b The “Cluster 9” group represents Cluster 9 in snRNA-seq dataset, and the “Other clusters” group represents cluster 0-8 in snRNA-seq dataset. c The “Normal” group represents untreated neural cells, the “Control” group represents neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group represents neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . d The “Apoptosis” group represents apoptosis cluster in snRNA-seq dataset, and the “Other clusters” group represents stable state, H 2 O 2 sensitive, SAOs sensitive, and others clusters in snRNA-seq dataset. e – i The “Normal” group represents untreated neural cells, the “Control” group comprises neural cells exposed to 200 μM H 2 O 2 , and the “SAOs” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 . The “mSIRK” group consists of neural cells treated with SAOs following exposure to 200 μM H 2 O 2 and 10 μM mSIRK (ERK1/2 activators). Source data are provided as a file.

    Article Snippet: Antibody include C-myc (18583S, Cell Signaling Technology), Nestin (ab105389, Abcam), Sox2 (3579S, Cell Signaling Technology), Caspase 9 (9508S, Cell Signaling Technology), Caspase 3(9662S, Cell Signaling Technology), p44/42 MAPK (Erk1/2) (4695S, Cell Signaling Technology), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (4370S, Cell Signaling Technology), p38 MAPK (8690S, Cell Signaling Technology), Phospho-p38 MAPK (Thr180/Tyr182) (4511S, Cell Signaling Technology), SAPK/JNK Antibody (9252S, Cell Signaling Technology), Phospho-SAPK/JNK (Thr183/Tyr185) (9255S, Cell Signaling Technology), GAPDH (ab125247, Abcam;), Anti-mouse IgG, HRP-linked Antibody (7076S, Cell Signaling Technology), Anti-rabbit IgG, HRP-linked Antibody (7074S, Cell Signaling Technology).

    Techniques: RNA Sequencing Assay, TUNEL Assay, Lactate Dehydrogenase Assay, Western Blot, Control