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


Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    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
    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, 2024-05
    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 p38 mapk  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc phospho p38 mapk
    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. <t>MAPK:</t> Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Phospho P38 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/phospho p38 mapk/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 mapk - by Bioz Stars, 2024-05
    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:

    anti phospho p38 t180 y182  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc anti phospho p38 t180 y182
    (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of <t>phospho-p38</t> levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .
    Anti Phospho P38 T180 Y182, 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/anti phospho p38 t180 y182/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 t180 y182 - by Bioz Stars, 2024-05
    86/100 stars

    Images

    1) Product Images from "Non-cell-autonomous regulation of mTORC2 by Hedgehog signaling maintains lipid homeostasis"

    Article Title: Non-cell-autonomous regulation of mTORC2 by Hedgehog signaling maintains lipid homeostasis

    Journal: bioRxiv

    doi: 10.1101/2024.05.06.592795

    (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of phospho-p38 levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .
    Figure Legend Snippet: (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of phospho-p38 levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .

    Techniques Used: Staining, Mutagenesis, Expressing, Western Blot

    anti phospho p38 t180 y182  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc anti phospho p38 t180 y182
    (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of <t>phospho-p38</t> levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .
    Anti Phospho P38 T180 Y182, 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/anti phospho p38 t180 y182/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 t180 y182 - by Bioz Stars, 2024-05
    86/100 stars

    Images

    1) Product Images from "Non-cell-autonomous regulation of mTORC2 by Hedgehog signaling maintains lipid homeostasis"

    Article Title: Non-cell-autonomous regulation of mTORC2 by Hedgehog signaling maintains lipid homeostasis

    Journal: bioRxiv

    doi: 10.1101/2024.05.06.592795

    (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of phospho-p38 levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .
    Figure Legend Snippet: (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of phospho-p38 levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .

    Techniques Used: Staining, Mutagenesis, Expressing, Western Blot

    rabbit anti phospho p38 mapk  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc rabbit anti phospho p38 mapk
    A, Bubble plot representing the enrichment analysis of 530 DEGs performed in SILAC Phosphoproteomics data. Color and x axis represent minus logarithms of p Value. The size represents numbers of genes enriched in the indicated data. B , Physical interactions, obtained by GeneMANIA, highlight Ezrin and EGFR binding. C , Volcano plot of DEGs, with up-regulated EGFR and down-regulated MAP2 and ERBB2 (no threshold on Log2FC and 0.05 threshold on -Log10FDR). Legend: red dot, up-regulated gene; blue dot, down-regulated gene; grey dot, not significant gene. D, Co-IP data for Ezrin-EGFR interaction. For the co-IP analyses, was used Ezrin antibody, conjugated with beads, and immunoblotted with EGFR antibody for WT and EZR −/− (left) and HeLa EZR T567D and EZR T567A (right) HeLa cells, respectively. Schematic representation of HeLa EZR T567D and EZR T567A co-IP (bottom). E , Confocal microscopy images showing EGFR (green) and EZR (red) co-localization on the membrane in HeLa WT cells (left) and magnified views of the regions are provided (right). Scale bar 10 µm (magnification 1 µm). Representative plots of co-localization profiles on the membrane between EGFR (green) and EZR (red). F , Immunofluorescent labelling images of EGFR in HeLa WT and EZR −/− cells, observed by confocal microscopy. Scale bar 10 µm. G , Immunoblots and calculated levels (bottom) of HER3, pY845 EGFR, EGFR, pT222 MK2, MK2, pT180/pY182 <t>p38</t> <t>MAPK</t> and <t>P38</t> <t>MAPK</t> in HeLa WT and EZR −/− cells. Data are expressed as mean of pY845EGFR/EGFR, pT222 MK2/MK2 and pT180/pY182 <t>p38</t> <t>MAPK/P38</t> MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: unpaired t-test for pY845 EGFR; Mann-Whitney test for HER3, EGFR, pT222 MK2, pT180/pY182 p38 MAPK.
    Rabbit Anti Phospho P38 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 anti phospho p38 mapk/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho p38 mapk - by Bioz Stars, 2024-05
    86/100 stars

    Images

    1) Product Images from "Ezrin defines TSC1 activation at endosomal compartments through EGFR-AKT signaling"

    Article Title: Ezrin defines TSC1 activation at endosomal compartments through EGFR-AKT signaling

    Journal: bioRxiv

    doi: 10.1101/2024.05.03.592332

    A, Bubble plot representing the enrichment analysis of 530 DEGs performed in SILAC Phosphoproteomics data. Color and x axis represent minus logarithms of p Value. The size represents numbers of genes enriched in the indicated data. B , Physical interactions, obtained by GeneMANIA, highlight Ezrin and EGFR binding. C , Volcano plot of DEGs, with up-regulated EGFR and down-regulated MAP2 and ERBB2 (no threshold on Log2FC and 0.05 threshold on -Log10FDR). Legend: red dot, up-regulated gene; blue dot, down-regulated gene; grey dot, not significant gene. D, Co-IP data for Ezrin-EGFR interaction. For the co-IP analyses, was used Ezrin antibody, conjugated with beads, and immunoblotted with EGFR antibody for WT and EZR −/− (left) and HeLa EZR T567D and EZR T567A (right) HeLa cells, respectively. Schematic representation of HeLa EZR T567D and EZR T567A co-IP (bottom). E , Confocal microscopy images showing EGFR (green) and EZR (red) co-localization on the membrane in HeLa WT cells (left) and magnified views of the regions are provided (right). Scale bar 10 µm (magnification 1 µm). Representative plots of co-localization profiles on the membrane between EGFR (green) and EZR (red). F , Immunofluorescent labelling images of EGFR in HeLa WT and EZR −/− cells, observed by confocal microscopy. Scale bar 10 µm. G , Immunoblots and calculated levels (bottom) of HER3, pY845 EGFR, EGFR, pT222 MK2, MK2, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells. Data are expressed as mean of pY845EGFR/EGFR, pT222 MK2/MK2 and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: unpaired t-test for pY845 EGFR; Mann-Whitney test for HER3, EGFR, pT222 MK2, pT180/pY182 p38 MAPK.
    Figure Legend Snippet: A, Bubble plot representing the enrichment analysis of 530 DEGs performed in SILAC Phosphoproteomics data. Color and x axis represent minus logarithms of p Value. The size represents numbers of genes enriched in the indicated data. B , Physical interactions, obtained by GeneMANIA, highlight Ezrin and EGFR binding. C , Volcano plot of DEGs, with up-regulated EGFR and down-regulated MAP2 and ERBB2 (no threshold on Log2FC and 0.05 threshold on -Log10FDR). Legend: red dot, up-regulated gene; blue dot, down-regulated gene; grey dot, not significant gene. D, Co-IP data for Ezrin-EGFR interaction. For the co-IP analyses, was used Ezrin antibody, conjugated with beads, and immunoblotted with EGFR antibody for WT and EZR −/− (left) and HeLa EZR T567D and EZR T567A (right) HeLa cells, respectively. Schematic representation of HeLa EZR T567D and EZR T567A co-IP (bottom). E , Confocal microscopy images showing EGFR (green) and EZR (red) co-localization on the membrane in HeLa WT cells (left) and magnified views of the regions are provided (right). Scale bar 10 µm (magnification 1 µm). Representative plots of co-localization profiles on the membrane between EGFR (green) and EZR (red). F , Immunofluorescent labelling images of EGFR in HeLa WT and EZR −/− cells, observed by confocal microscopy. Scale bar 10 µm. G , Immunoblots and calculated levels (bottom) of HER3, pY845 EGFR, EGFR, pT222 MK2, MK2, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells. Data are expressed as mean of pY845EGFR/EGFR, pT222 MK2/MK2 and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: unpaired t-test for pY845 EGFR; Mann-Whitney test for HER3, EGFR, pT222 MK2, pT180/pY182 p38 MAPK.

    Techniques Used: Binding Assay, Co-Immunoprecipitation Assay, Confocal Microscopy, Membrane, Western Blot, MANN-WHITNEY

    A , Live cell imaging and model for EGFR (green) translocation from the membrane to the endosomes in HeLa WT (top) and EZR −/− (bottom) cells without EGF stimulation (T0) and with a progressive EGF stimulation (from T10’’ to T60’’). White boxes are magnifications that depict EGFR protein migration. Scale bar 1 µm). Please refer to Video 1. B , IEM (anti-GFP immunolabelling) of cycloheximide treated HeLa WT, WT + EGF, EZR −/− and EZR −/− + EGF cells expressing EGFR-GFP. Endosomes containing EGFR is shown in green. Scale bar 200 nm. Quantitative analysis (right) of EGFR positive endosomes expressed as mean ± SEM. Statistical test: generalized Linear Model with Likelihood Ratio (Poisson Regression). C , immunoblots and calculated levels (bottom) of HER2, pY845 EGFR, EGFR, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells with (+) and without (-) EGF stimulation. Data are expressed as mean of pY845EGFR/EGFR and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: Unpaired t-test for HER2 WT, HER2 EZR −/− , pY845 EGFR EZR −/− , EGFR WT, EGFR EZR −/− , pT180/pY182 p38 MAPK WT, pT180/pY182 p38 MAPK EZR −/− ; unpaired t-test with Welch’s correction for pY845 EGFR WT. D , Representative immunoblots of EGFR in membrane (top) and endosomes (bottom) proteins in HeLa WT and EZR −/− with (+) and without (-) EGF stimulation. ZO-1 and EEA1 are used as membrane and endosomes extraction control, respectively. GAPDH are used as loading control.
    Figure Legend Snippet: A , Live cell imaging and model for EGFR (green) translocation from the membrane to the endosomes in HeLa WT (top) and EZR −/− (bottom) cells without EGF stimulation (T0) and with a progressive EGF stimulation (from T10’’ to T60’’). White boxes are magnifications that depict EGFR protein migration. Scale bar 1 µm). Please refer to Video 1. B , IEM (anti-GFP immunolabelling) of cycloheximide treated HeLa WT, WT + EGF, EZR −/− and EZR −/− + EGF cells expressing EGFR-GFP. Endosomes containing EGFR is shown in green. Scale bar 200 nm. Quantitative analysis (right) of EGFR positive endosomes expressed as mean ± SEM. Statistical test: generalized Linear Model with Likelihood Ratio (Poisson Regression). C , immunoblots and calculated levels (bottom) of HER2, pY845 EGFR, EGFR, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells with (+) and without (-) EGF stimulation. Data are expressed as mean of pY845EGFR/EGFR and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: Unpaired t-test for HER2 WT, HER2 EZR −/− , pY845 EGFR EZR −/− , EGFR WT, EGFR EZR −/− , pT180/pY182 p38 MAPK WT, pT180/pY182 p38 MAPK EZR −/− ; unpaired t-test with Welch’s correction for pY845 EGFR WT. D , Representative immunoblots of EGFR in membrane (top) and endosomes (bottom) proteins in HeLa WT and EZR −/− with (+) and without (-) EGF stimulation. ZO-1 and EEA1 are used as membrane and endosomes extraction control, respectively. GAPDH are used as loading control.

    Techniques Used: Live Cell Imaging, Translocation Assay, Membrane, Migration, Expressing, Western Blot, Extraction

    phospho p38 mapk thr180 tyr182 antibody  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 antibody
    Treatment of cells with Aβ led to increased phospho/total <t>p38</t> <t>MAPK</t> ratio, while blocking <t>p38</t> <t>MAPK</t> activity with SB203580 decreased p53 activity in A549 cells and increased ACh levels in the media of both cell lines. Cells (0.2 × 10) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight. The cells were then treated as indicated for 72 h with Aβ 1–40/42 or fragments (10 μM) ± SB203580 (SB, 20 μM), and then, the p38 MAPK assay ( A ) and Western blotting ( B ) were carried out (Materials and Methods). The p53 activity was measured in A549 cells ( C ). Cells transfected with either control or p53 siRNA were treated for 72 h with Aβ 1–40/42 or fragments ± SB203580; then, the levels of ACh ( D – F ) were measured as described in the Materials and Methods Section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to the control of each cell line ( A ) or to control in the absence of SB203580 ( C – F ) using the GraphPad 10.1.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks indicate a statistically significant difference from control. Absence of asterisks indicates no significance, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple comparison test, * p < 0.05, ** p < 0.01.
    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
    https://www.bioz.com/result/phospho p38 mapk thr180 tyr182 antibody/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 antibody - by Bioz Stars, 2024-05
    86/100 stars

    Images

    1) Product Images from "Amyloid Beta Leads to Decreased Acetylcholine Levels and Non-Small Cell Lung Cancer Cell Survival via a Mechanism That Involves p38 Mitogen-Activated Protein Kinase and Protein Kinase C in a p53-Dependent and -Independent Manner"

    Article Title: Amyloid Beta Leads to Decreased Acetylcholine Levels and Non-Small Cell Lung Cancer Cell Survival via a Mechanism That Involves p38 Mitogen-Activated Protein Kinase and Protein Kinase C in a p53-Dependent and -Independent Manner

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms25095033

    Treatment of cells with Aβ led to increased phospho/total p38 MAPK ratio, while blocking p38 MAPK activity with SB203580 decreased p53 activity in A549 cells and increased ACh levels in the media of both cell lines. Cells (0.2 × 10) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight. The cells were then treated as indicated for 72 h with Aβ 1–40/42 or fragments (10 μM) ± SB203580 (SB, 20 μM), and then, the p38 MAPK assay ( A ) and Western blotting ( B ) were carried out (Materials and Methods). The p53 activity was measured in A549 cells ( C ). Cells transfected with either control or p53 siRNA were treated for 72 h with Aβ 1–40/42 or fragments ± SB203580; then, the levels of ACh ( D – F ) were measured as described in the Materials and Methods Section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to the control of each cell line ( A ) or to control in the absence of SB203580 ( C – F ) using the GraphPad 10.1.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks indicate a statistically significant difference from control. Absence of asterisks indicates no significance, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple comparison test, * p < 0.05, ** p < 0.01.
    Figure Legend Snippet: Treatment of cells with Aβ led to increased phospho/total p38 MAPK ratio, while blocking p38 MAPK activity with SB203580 decreased p53 activity in A549 cells and increased ACh levels in the media of both cell lines. Cells (0.2 × 10) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight. The cells were then treated as indicated for 72 h with Aβ 1–40/42 or fragments (10 μM) ± SB203580 (SB, 20 μM), and then, the p38 MAPK assay ( A ) and Western blotting ( B ) were carried out (Materials and Methods). The p53 activity was measured in A549 cells ( C ). Cells transfected with either control or p53 siRNA were treated for 72 h with Aβ 1–40/42 or fragments ± SB203580; then, the levels of ACh ( D – F ) were measured as described in the Materials and Methods Section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to the control of each cell line ( A ) or to control in the absence of SB203580 ( C – F ) using the GraphPad 10.1.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks indicate a statistically significant difference from control. Absence of asterisks indicates no significance, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple comparison test, * p < 0.05, ** p < 0.01.

    Techniques Used: Blocking Assay, Activity Assay, Western Blot, Transfection, Software, MANN-WHITNEY, Comparison

    Incubation of cells with ACh reduced the activity of p53 in A549 cells, diminished the activity of p38 MAPK, and enhanced the PKC activity in both A549 and H1299 cells, while co-incubation with Aβ reduced those effects. Cells (0.2 × 10 5 ) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight (Control, 0 hour). The cells were then treated as indicated with ACh (100 nM) and in combination with Aβ 1–40/42 or fragments (10 μM). The p53 activity ( A ) in A549 cells and the activity of p38 MAPK ( B , C ) and PKC ( D , E ) were measured as described in the Materials and Methods Section. The data were expressed as the percentage of control by expressing each point relative to the control (set to 100%). The data were then plotted as a function of time using the GraphPad Prism 10.1.1 software. Data were expressed as the mean ± S.D. of three independent experiments, each carried out in triplicate.
    Figure Legend Snippet: Incubation of cells with ACh reduced the activity of p53 in A549 cells, diminished the activity of p38 MAPK, and enhanced the PKC activity in both A549 and H1299 cells, while co-incubation with Aβ reduced those effects. Cells (0.2 × 10 5 ) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight (Control, 0 hour). The cells were then treated as indicated with ACh (100 nM) and in combination with Aβ 1–40/42 or fragments (10 μM). The p53 activity ( A ) in A549 cells and the activity of p38 MAPK ( B , C ) and PKC ( D , E ) were measured as described in the Materials and Methods Section. The data were expressed as the percentage of control by expressing each point relative to the control (set to 100%). The data were then plotted as a function of time using the GraphPad Prism 10.1.1 software. Data were expressed as the mean ± S.D. of three independent experiments, each carried out in triplicate.

    Techniques Used: Incubation, Activity Assay, Expressing, Software

    Representation of the main hypothesis and findings of this study. Aβ decreases the levels of ACh in the media via the activation of p53/AChE and p38 MAPK and/or via blocking the activity of PKC, leading to decreased cell survival. In this model, ACh reverses the effects of Aβ.
    Figure Legend Snippet: Representation of the main hypothesis and findings of this study. Aβ decreases the levels of ACh in the media via the activation of p53/AChE and p38 MAPK and/or via blocking the activity of PKC, leading to decreased cell survival. In this model, ACh reverses the effects of Aβ.

    Techniques Used: Activation Assay, Blocking Assay, Activity Assay

    anti phospho p38 mitogen  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc anti phospho p38 mitogen
    Anti Phospho P38 Mitogen, 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/anti phospho p38 mitogen/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 mitogen - by Bioz Stars, 2024-05
    86/100 stars

    Images

    anti phospho p38 mitogen  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc anti phospho p38 mitogen
    CTSK knockout improved the stress-related harmful alterations of the targeted protein levels in the FeCl 3 -treated arteries after 21 days of the stress protocol. A – C Total protein was isolated from the thrombotic arterial tissues of the stressed CTSK + / + and stressed CTSK −/− mice and applied to western blotting. Representative images and quantitative data show the levels of CTSK, c-Notch1, Hes1, p16 IN4A , gp91phox, c-caspase8, ATR1, and <t>p-p38MAPK</t> proteins in both groups. Results are mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. corresponding non-stressed or stressed CTSS + / + mice by one-way ANOVA and Tukey’s post hoc test
    Anti Phospho P38 Mitogen, 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/anti phospho p38 mitogen/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 mitogen - by Bioz Stars, 2024-05
    86/100 stars

    Images

    1) Product Images from "Cathepsin K deficiency prevented stress-related thrombosis in a mouse FeCl 3 model"

    Article Title: Cathepsin K deficiency prevented stress-related thrombosis in a mouse FeCl 3 model

    Journal: Cellular and Molecular Life Sciences: CMLS

    doi: 10.1007/s00018-024-05240-0

    CTSK knockout improved the stress-related harmful alterations of the targeted protein levels in the FeCl 3 -treated arteries after 21 days of the stress protocol. A – C Total protein was isolated from the thrombotic arterial tissues of the stressed CTSK + / + and stressed CTSK −/− mice and applied to western blotting. Representative images and quantitative data show the levels of CTSK, c-Notch1, Hes1, p16 IN4A , gp91phox, c-caspase8, ATR1, and p-p38MAPK proteins in both groups. Results are mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. corresponding non-stressed or stressed CTSS + / + mice by one-way ANOVA and Tukey’s post hoc test
    Figure Legend Snippet: CTSK knockout improved the stress-related harmful alterations of the targeted protein levels in the FeCl 3 -treated arteries after 21 days of the stress protocol. A – C Total protein was isolated from the thrombotic arterial tissues of the stressed CTSK + / + and stressed CTSK −/− mice and applied to western blotting. Representative images and quantitative data show the levels of CTSK, c-Notch1, Hes1, p16 IN4A , gp91phox, c-caspase8, ATR1, and p-p38MAPK proteins in both groups. Results are mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. corresponding non-stressed or stressed CTSS + / + mice by one-way ANOVA and Tukey’s post hoc test

    Techniques Used: Knock-Out, Isolation, Western Blot

    rabbit anti phospho p38 antibody  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc rabbit anti phospho p38 antibody
    Western blotting analysis of the relative expression of MAPK signaling pathway components in the retina in the different intervention groups. The rats were subjected to IOP elevation, and then received the intravitreal injection of DMSO, the CK2 inhibitor TBB, the CK2 inhibitor DMAT, and the macrophage activator ZYM. Rats that were not subjected to IOP elevation and were intravitreally injected with DMSO served as intact controls. (A) The relative protein expression levels of <t>p38</t> and p-p38 were examined by western blotting. (B) The relative protein expression levels of Erk p44/p42 and p-p44/p42 were detected by western blotting. The results are presented as histograms after normalization to β-actin or GAPDH. The data are presented as mean ± SD ( n = 5). * P < 0.05 (one-way analysis of variance among multiple groups followed by post hoc Bonferroni test). CK2: Casein kinase-2; DMAT: 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole; DMSO: dimethylsulfoxide; Erk: extracellular-signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IOP: intraocular pressure; MAPK: mitogen-activated protein kinase; RGCs: retinal ganglion cells; TBB: 4,5,6,7-tetrabromo-2-azabenzimidazole; ZYM: zymosan.
    Rabbit Anti Phospho P38 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
    https://www.bioz.com/result/rabbit anti phospho p38 antibody/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho p38 antibody - by Bioz Stars, 2024-05
    86/100 stars

    Images

    1) Product Images from "Casein kinase-2 inhibition promotes retinal ganglion cell survival after acute intraocular pressure elevation"

    Article Title: Casein kinase-2 inhibition promotes retinal ganglion cell survival after acute intraocular pressure elevation

    Journal: Neural Regeneration Research

    doi: 10.4103/1673-5374.385310

    Western blotting analysis of the relative expression of MAPK signaling pathway components in the retina in the different intervention groups. The rats were subjected to IOP elevation, and then received the intravitreal injection of DMSO, the CK2 inhibitor TBB, the CK2 inhibitor DMAT, and the macrophage activator ZYM. Rats that were not subjected to IOP elevation and were intravitreally injected with DMSO served as intact controls. (A) The relative protein expression levels of p38 and p-p38 were examined by western blotting. (B) The relative protein expression levels of Erk p44/p42 and p-p44/p42 were detected by western blotting. The results are presented as histograms after normalization to β-actin or GAPDH. The data are presented as mean ± SD ( n = 5). * P < 0.05 (one-way analysis of variance among multiple groups followed by post hoc Bonferroni test). CK2: Casein kinase-2; DMAT: 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole; DMSO: dimethylsulfoxide; Erk: extracellular-signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IOP: intraocular pressure; MAPK: mitogen-activated protein kinase; RGCs: retinal ganglion cells; TBB: 4,5,6,7-tetrabromo-2-azabenzimidazole; ZYM: zymosan.
    Figure Legend Snippet: Western blotting analysis of the relative expression of MAPK signaling pathway components in the retina in the different intervention groups. The rats were subjected to IOP elevation, and then received the intravitreal injection of DMSO, the CK2 inhibitor TBB, the CK2 inhibitor DMAT, and the macrophage activator ZYM. Rats that were not subjected to IOP elevation and were intravitreally injected with DMSO served as intact controls. (A) The relative protein expression levels of p38 and p-p38 were examined by western blotting. (B) The relative protein expression levels of Erk p44/p42 and p-p44/p42 were detected by western blotting. The results are presented as histograms after normalization to β-actin or GAPDH. The data are presented as mean ± SD ( n = 5). * P < 0.05 (one-way analysis of variance among multiple groups followed by post hoc Bonferroni test). CK2: Casein kinase-2; DMAT: 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole; DMSO: dimethylsulfoxide; Erk: extracellular-signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IOP: intraocular pressure; MAPK: mitogen-activated protein kinase; RGCs: retinal ganglion cells; TBB: 4,5,6,7-tetrabromo-2-azabenzimidazole; ZYM: zymosan.

    Techniques Used: Western Blot, Expressing, Injection

    phospho p38  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Cell Signaling Technology Inc phospho p38
    LPA 1 antagonist attenuates NF-κB upregulation and MAPK activation but enhances PI3K/Akt activation after pMCAO challenge. Mice were challenged with pMCAO. AM152 (5 mg/kg, p.o. ) was then administered at one hour after occlusion. (A) Effects of AM152 on NF-κB expression in activated microglia were determined by NF-κB(p65)/Iba1 double immunofluorescence at 1 day after pMCAO challenge. Representative images of NF-κB(p65) /Iba1-double immunopositive cells in ischemic core regions and quantification are shown. Scale bar, 50 μm. n=4 mice per group. *** p <0.001 versus sham. ### p <0.001 versus vehicle-administered pMCAO group. (B) Effects of AM152 on phosphorylation of MAPKs (ERK1/2, <t>p38,</t> and JNK) and PI3K/Akt were determined by Western blot analysis at 1 day after pMCAO challenge. Representative Western blots and quantification are shown. n=3 mice per group. * p <0.05, ** p <0.01, and *** p <0.001 versus sham. # p <0.05, ## p <0.01 and ### p <0.001 versus vehicle-administered pMCAO group.
    Phospho P38, 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/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 - by Bioz Stars, 2024-05
    86/100 stars

    Images

    1) Product Images from "Lysophosphatidic Acid Receptor 1 Plays a Pathogenic Role in Permanent Brain Ischemic Stroke by Modulating Neuroinflammatory Responses"

    Article Title: Lysophosphatidic Acid Receptor 1 Plays a Pathogenic Role in Permanent Brain Ischemic Stroke by Modulating Neuroinflammatory Responses

    Journal: Biomolecules & Therapeutics

    doi: 10.4062/biomolther.2024.052

    LPA 1 antagonist attenuates NF-κB upregulation and MAPK activation but enhances PI3K/Akt activation after pMCAO challenge. Mice were challenged with pMCAO. AM152 (5 mg/kg, p.o. ) was then administered at one hour after occlusion. (A) Effects of AM152 on NF-κB expression in activated microglia were determined by NF-κB(p65)/Iba1 double immunofluorescence at 1 day after pMCAO challenge. Representative images of NF-κB(p65) /Iba1-double immunopositive cells in ischemic core regions and quantification are shown. Scale bar, 50 μm. n=4 mice per group. *** p <0.001 versus sham. ### p <0.001 versus vehicle-administered pMCAO group. (B) Effects of AM152 on phosphorylation of MAPKs (ERK1/2, p38, and JNK) and PI3K/Akt were determined by Western blot analysis at 1 day after pMCAO challenge. Representative Western blots and quantification are shown. n=3 mice per group. * p <0.05, ** p <0.01, and *** p <0.001 versus sham. # p <0.05, ## p <0.01 and ### p <0.001 versus vehicle-administered pMCAO group.
    Figure Legend Snippet: LPA 1 antagonist attenuates NF-κB upregulation and MAPK activation but enhances PI3K/Akt activation after pMCAO challenge. Mice were challenged with pMCAO. AM152 (5 mg/kg, p.o. ) was then administered at one hour after occlusion. (A) Effects of AM152 on NF-κB expression in activated microglia were determined by NF-κB(p65)/Iba1 double immunofluorescence at 1 day after pMCAO challenge. Representative images of NF-κB(p65) /Iba1-double immunopositive cells in ischemic core regions and quantification are shown. Scale bar, 50 μm. n=4 mice per group. *** p <0.001 versus sham. ### p <0.001 versus vehicle-administered pMCAO group. (B) Effects of AM152 on phosphorylation of MAPKs (ERK1/2, p38, and JNK) and PI3K/Akt were determined by Western blot analysis at 1 day after pMCAO challenge. Representative Western blots and quantification are shown. n=3 mice per group. * p <0.05, ** p <0.01, and *** p <0.001 versus sham. # p <0.05, ## p <0.01 and ### p <0.001 versus vehicle-administered pMCAO group.

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

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    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
    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, 2024-05
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc phospho p38 mapk
    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. <t>MAPK:</t> Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Phospho P38 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/phospho p38 mapk/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 mapk - by Bioz Stars, 2024-05
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc anti phospho p38 t180 y182
    (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of <t>phospho-p38</t> levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .
    Anti Phospho P38 T180 Y182, 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/anti phospho p38 t180 y182/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 t180 y182 - by Bioz Stars, 2024-05
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc rabbit anti phospho p38 mapk
    A, Bubble plot representing the enrichment analysis of 530 DEGs performed in SILAC Phosphoproteomics data. Color and x axis represent minus logarithms of p Value. The size represents numbers of genes enriched in the indicated data. B , Physical interactions, obtained by GeneMANIA, highlight Ezrin and EGFR binding. C , Volcano plot of DEGs, with up-regulated EGFR and down-regulated MAP2 and ERBB2 (no threshold on Log2FC and 0.05 threshold on -Log10FDR). Legend: red dot, up-regulated gene; blue dot, down-regulated gene; grey dot, not significant gene. D, Co-IP data for Ezrin-EGFR interaction. For the co-IP analyses, was used Ezrin antibody, conjugated with beads, and immunoblotted with EGFR antibody for WT and EZR −/− (left) and HeLa EZR T567D and EZR T567A (right) HeLa cells, respectively. Schematic representation of HeLa EZR T567D and EZR T567A co-IP (bottom). E , Confocal microscopy images showing EGFR (green) and EZR (red) co-localization on the membrane in HeLa WT cells (left) and magnified views of the regions are provided (right). Scale bar 10 µm (magnification 1 µm). Representative plots of co-localization profiles on the membrane between EGFR (green) and EZR (red). F , Immunofluorescent labelling images of EGFR in HeLa WT and EZR −/− cells, observed by confocal microscopy. Scale bar 10 µm. G , Immunoblots and calculated levels (bottom) of HER3, pY845 EGFR, EGFR, pT222 MK2, MK2, pT180/pY182 <t>p38</t> <t>MAPK</t> and <t>P38</t> <t>MAPK</t> in HeLa WT and EZR −/− cells. Data are expressed as mean of pY845EGFR/EGFR, pT222 MK2/MK2 and pT180/pY182 <t>p38</t> <t>MAPK/P38</t> MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: unpaired t-test for pY845 EGFR; Mann-Whitney test for HER3, EGFR, pT222 MK2, pT180/pY182 p38 MAPK.
    Rabbit Anti Phospho P38 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 anti phospho p38 mapk/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho p38 mapk - by Bioz Stars, 2024-05
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 antibody
    Treatment of cells with Aβ led to increased phospho/total <t>p38</t> <t>MAPK</t> ratio, while blocking <t>p38</t> <t>MAPK</t> activity with SB203580 decreased p53 activity in A549 cells and increased ACh levels in the media of both cell lines. Cells (0.2 × 10) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight. The cells were then treated as indicated for 72 h with Aβ 1–40/42 or fragments (10 μM) ± SB203580 (SB, 20 μM), and then, the p38 MAPK assay ( A ) and Western blotting ( B ) were carried out (Materials and Methods). The p53 activity was measured in A549 cells ( C ). Cells transfected with either control or p53 siRNA were treated for 72 h with Aβ 1–40/42 or fragments ± SB203580; then, the levels of ACh ( D – F ) were measured as described in the Materials and Methods Section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to the control of each cell line ( A ) or to control in the absence of SB203580 ( C – F ) using the GraphPad 10.1.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks indicate a statistically significant difference from control. Absence of asterisks indicates no significance, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple comparison test, * p < 0.05, ** p < 0.01.
    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
    https://www.bioz.com/result/phospho p38 mapk thr180 tyr182 antibody/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 antibody - by Bioz Stars, 2024-05
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc anti phospho p38 mitogen
    Treatment of cells with Aβ led to increased phospho/total <t>p38</t> <t>MAPK</t> ratio, while blocking <t>p38</t> <t>MAPK</t> activity with SB203580 decreased p53 activity in A549 cells and increased ACh levels in the media of both cell lines. Cells (0.2 × 10) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight. The cells were then treated as indicated for 72 h with Aβ 1–40/42 or fragments (10 μM) ± SB203580 (SB, 20 μM), and then, the p38 MAPK assay ( A ) and Western blotting ( B ) were carried out (Materials and Methods). The p53 activity was measured in A549 cells ( C ). Cells transfected with either control or p53 siRNA were treated for 72 h with Aβ 1–40/42 or fragments ± SB203580; then, the levels of ACh ( D – F ) were measured as described in the Materials and Methods Section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to the control of each cell line ( A ) or to control in the absence of SB203580 ( C – F ) using the GraphPad 10.1.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks indicate a statistically significant difference from control. Absence of asterisks indicates no significance, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple comparison test, * p < 0.05, ** p < 0.01.
    Anti Phospho P38 Mitogen, 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/anti phospho p38 mitogen/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho p38 mitogen - by Bioz Stars, 2024-05
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc rabbit anti phospho p38 antibody
    Western blotting analysis of the relative expression of MAPK signaling pathway components in the retina in the different intervention groups. The rats were subjected to IOP elevation, and then received the intravitreal injection of DMSO, the CK2 inhibitor TBB, the CK2 inhibitor DMAT, and the macrophage activator ZYM. Rats that were not subjected to IOP elevation and were intravitreally injected with DMSO served as intact controls. (A) The relative protein expression levels of <t>p38</t> and p-p38 were examined by western blotting. (B) The relative protein expression levels of Erk p44/p42 and p-p44/p42 were detected by western blotting. The results are presented as histograms after normalization to β-actin or GAPDH. The data are presented as mean ± SD ( n = 5). * P < 0.05 (one-way analysis of variance among multiple groups followed by post hoc Bonferroni test). CK2: Casein kinase-2; DMAT: 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole; DMSO: dimethylsulfoxide; Erk: extracellular-signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IOP: intraocular pressure; MAPK: mitogen-activated protein kinase; RGCs: retinal ganglion cells; TBB: 4,5,6,7-tetrabromo-2-azabenzimidazole; ZYM: zymosan.
    Rabbit Anti Phospho P38 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
    https://www.bioz.com/result/rabbit anti phospho p38 antibody/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti phospho p38 antibody - by Bioz Stars, 2024-05
    86/100 stars
      Buy from Supplier

    86
    Cell Signaling Technology Inc phospho p38
    LPA 1 antagonist attenuates NF-κB upregulation and MAPK activation but enhances PI3K/Akt activation after pMCAO challenge. Mice were challenged with pMCAO. AM152 (5 mg/kg, p.o. ) was then administered at one hour after occlusion. (A) Effects of AM152 on NF-κB expression in activated microglia were determined by NF-κB(p65)/Iba1 double immunofluorescence at 1 day after pMCAO challenge. Representative images of NF-κB(p65) /Iba1-double immunopositive cells in ischemic core regions and quantification are shown. Scale bar, 50 μm. n=4 mice per group. *** p <0.001 versus sham. ### p <0.001 versus vehicle-administered pMCAO group. (B) Effects of AM152 on phosphorylation of MAPKs (ERK1/2, <t>p38,</t> and JNK) and PI3K/Akt were determined by Western blot analysis at 1 day after pMCAO challenge. Representative Western blots and quantification are shown. n=3 mice per group. * p <0.05, ** p <0.01, and *** p <0.001 versus sham. # p <0.05, ## p <0.01 and ### p <0.001 versus vehicle-administered pMCAO group.
    Phospho P38, 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/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho p38 - by Bioz Stars, 2024-05
    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:

    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 membrane were incubated with blocking buffer (5% nonfat milk in TBST) for 90 minutes at room temperature, followed by incubation with primary antibodies overnight at 4°C: beta-actin monoclonal antibody (1:1000, Proteintech, Rosemont, IL, USA, Cat# 66009-1-Ig, RRID: AB_2687938), p38 MAPK polyclonal antibody (1:1000, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), ferritin light chain (FTL) polyclonal antibody (1:1000, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673), and SAT1 polyclonal antibody (1:1000, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK (1:1000, Thr180/Tyr182 Rabbit mAb, Cell Signaling Technology, Danvers, MA, USA, Cat# 4511); recombinant anti-glutathione peroxidase 4 (GPx4) antibody (1:1000, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 polyclonal antibody (1:1000, Thermo Fisher Scientific, 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 membrane were incubated with blocking buffer (5% nonfat milk in TBST) for 90 minutes at room temperature, followed by incubation with primary antibodies overnight at 4°C: beta-actin monoclonal antibody (1:1000, Proteintech, Rosemont, IL, USA, Cat# 66009-1-Ig, RRID: AB_2687938), p38 MAPK polyclonal antibody (1:1000, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), ferritin light chain (FTL) polyclonal antibody (1:1000, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673), and SAT1 polyclonal antibody (1:1000, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK (1:1000, Thr180/Tyr182 Rabbit mAb, Cell Signaling Technology, Danvers, MA, USA, Cat# 4511); recombinant anti-glutathione peroxidase 4 (GPx4) antibody (1:1000, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 polyclonal antibody (1:1000, Thermo Fisher Scientific, 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 membrane were incubated with blocking buffer (5% nonfat milk in TBST) for 90 minutes at room temperature, followed by incubation with primary antibodies overnight at 4°C: beta-actin monoclonal antibody (1:1000, Proteintech, Rosemont, IL, USA, Cat# 66009-1-Ig, RRID: AB_2687938), p38 MAPK polyclonal antibody (1:1000, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), ferritin light chain (FTL) polyclonal antibody (1:1000, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673), and SAT1 polyclonal antibody (1:1000, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK (1:1000, Thr180/Tyr182 Rabbit mAb, Cell Signaling Technology, Danvers, MA, USA, Cat# 4511); recombinant anti-glutathione peroxidase 4 (GPx4) antibody (1:1000, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 polyclonal antibody (1:1000, Thermo Fisher Scientific, 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 membrane were incubated with blocking buffer (5% nonfat milk in TBST) for 90 minutes at room temperature, followed by incubation with primary antibodies overnight at 4°C: beta-actin monoclonal antibody (1:1000, Proteintech, Rosemont, IL, USA, Cat# 66009-1-Ig, RRID: AB_2687938), p38 MAPK polyclonal antibody (1:1000, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), ferritin light chain (FTL) polyclonal antibody (1:1000, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673), and SAT1 polyclonal antibody (1:1000, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK (1:1000, Thr180/Tyr182 Rabbit mAb, Cell Signaling Technology, Danvers, MA, USA, Cat# 4511); recombinant anti-glutathione peroxidase 4 (GPx4) antibody (1:1000, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 polyclonal antibody (1:1000, Thermo Fisher Scientific, 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 membrane were incubated with blocking buffer (5% nonfat milk in TBST) for 90 minutes at room temperature, followed by incubation with primary antibodies overnight at 4°C: beta-actin monoclonal antibody (1:1000, Proteintech, Rosemont, IL, USA, Cat# 66009-1-Ig, RRID: AB_2687938), p38 MAPK polyclonal antibody (1:1000, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), ferritin light chain (FTL) polyclonal antibody (1:1000, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673), and SAT1 polyclonal antibody (1:1000, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK (1:1000, Thr180/Tyr182 Rabbit mAb, Cell Signaling Technology, Danvers, MA, USA, Cat# 4511); recombinant anti-glutathione peroxidase 4 (GPx4) antibody (1:1000, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 polyclonal antibody (1:1000, Thermo Fisher Scientific, 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 membrane were incubated with blocking buffer (5% nonfat milk in TBST) for 90 minutes at room temperature, followed by incubation with primary antibodies overnight at 4°C: beta-actin monoclonal antibody (1:1000, Proteintech, Rosemont, IL, USA, Cat# 66009-1-Ig, RRID: AB_2687938), p38 MAPK polyclonal antibody (1:1000, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), ferritin light chain (FTL) polyclonal antibody (1:1000, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673), and SAT1 polyclonal antibody (1:1000, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK (1:1000, Thr180/Tyr182 Rabbit mAb, Cell Signaling Technology, Danvers, MA, USA, Cat# 4511); recombinant anti-glutathione peroxidase 4 (GPx4) antibody (1:1000, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 polyclonal antibody (1:1000, Thermo Fisher Scientific, 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 membrane were incubated with blocking buffer (5% nonfat milk in TBST) for 90 minutes at room temperature, followed by incubation with primary antibodies overnight at 4°C: beta-actin monoclonal antibody (1:1000, Proteintech, Rosemont, IL, USA, Cat# 66009-1-Ig, RRID: AB_2687938), p38 MAPK polyclonal antibody (1:1000, Proteintech, Cat# 14064-1-AP, RRID: AB_2878007), ferritin light chain (FTL) polyclonal antibody (1:1000, Proteintech, Cat# 10727-1-AP, RRID: AB_2278673), and SAT1 polyclonal antibody (1:1000, Proteintech, Cat# 10708-1-AP, RRID: AB_2877739); phospho-p38 MAPK (1:1000, Thr180/Tyr182 Rabbit mAb, Cell Signaling Technology, Danvers, MA, USA, Cat# 4511); recombinant anti-glutathione peroxidase 4 (GPx4) antibody (1:1000, Abcam, Cat# ab125066, RRID: AB_10973901); and SLC7A11 polyclonal antibody (1:1000, Thermo Fisher Scientific, PA1-16893).

    Techniques:

    (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of phospho-p38 levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .

    Journal: bioRxiv

    Article Title: Non-cell-autonomous regulation of mTORC2 by Hedgehog signaling maintains lipid homeostasis

    doi: 10.1101/2024.05.06.592795

    Figure Lengend Snippet: (A) A Venn diagram showing that the rict-1/grd co-regulated genes are enriched for pmk-1 -dependent genes (hypergeometric P value reported ). (B) Quantification of lipid levels using Nile Red staining of the indicated wild-type and mutant strains. Data are representated as the mean ± SD (****, P <0.0001, ***, P <0.001, **, P <0.01, one-way ANOVA). (C) A heatmap of the 1,632 differentially expressed genes in grd mutants showing differential expression values (log2 fold change relative to wild-type) for the indicated mutant strains. Each row represents a gene organized from most up-regulated to most down-regulated in grd and R 2 values are shown to compare grd to grd; tir-1 and rict-1 , as well as rict-1 to rict-1; tir-1 . Western blot analyses of phospho-p38 levels for (D) C. elegans lysates prepared from wild-type, rict-1(mg360) , grd , or nsy-1(ums8) mutants ( ums8 is a gain-of-function allele) and (E) AML12 hepatocytes treated with SAG for increasing amounts of time. The pPMK-1/PMK-1 ratio is plotted for the C. elegans data. (F) A model of how Hh governs intestinal metabolism through the dual regulation of mTORC2 and p38 signaling in C. elegans .

    Article Snippet: Protein samples (∼50 μg) were resolved by SDS-PAGE, transferred to a PVDF membrane, and probed with either anti-FLAG (M2, Sigma, F1804), anti-HA (3F10, Sigma), anti-Actin (ab3280, Abcam), anti-pAkt S473 (D9E, Cell Signaling Technology), anti-pAkt T308 (D25E6, Cell Signaling Technology), anti-Akt (Cell Signaling Technology), anti-GLI1 (C68H3, Cell Signaling Technology), anti-pFoxO1/3a/4 T24/T32/T28 (4G6, Cell Signaling Technology), anti-FoxO1 (C29H4, Cell Signaling Technology), anti-Rictor (Cell Signaling Technology), anti-GFP (GF28R, Invitrogen), anti-phospho-p38 T180/Y182 for C. elegans (3D7, Cell Signaling Technology), anti-phospho-p38 T180/Y182 for mammalian samples (9211, Cell Signaling Technology), anti-p38 for C. elegans (gift from Dr. Read Pukkila-Worley), or anti-p38 for mammalian samples (9212, Cell Signaling Technology) antibodies.

    Techniques: Staining, Mutagenesis, Expressing, Western Blot

    A, Bubble plot representing the enrichment analysis of 530 DEGs performed in SILAC Phosphoproteomics data. Color and x axis represent minus logarithms of p Value. The size represents numbers of genes enriched in the indicated data. B , Physical interactions, obtained by GeneMANIA, highlight Ezrin and EGFR binding. C , Volcano plot of DEGs, with up-regulated EGFR and down-regulated MAP2 and ERBB2 (no threshold on Log2FC and 0.05 threshold on -Log10FDR). Legend: red dot, up-regulated gene; blue dot, down-regulated gene; grey dot, not significant gene. D, Co-IP data for Ezrin-EGFR interaction. For the co-IP analyses, was used Ezrin antibody, conjugated with beads, and immunoblotted with EGFR antibody for WT and EZR −/− (left) and HeLa EZR T567D and EZR T567A (right) HeLa cells, respectively. Schematic representation of HeLa EZR T567D and EZR T567A co-IP (bottom). E , Confocal microscopy images showing EGFR (green) and EZR (red) co-localization on the membrane in HeLa WT cells (left) and magnified views of the regions are provided (right). Scale bar 10 µm (magnification 1 µm). Representative plots of co-localization profiles on the membrane between EGFR (green) and EZR (red). F , Immunofluorescent labelling images of EGFR in HeLa WT and EZR −/− cells, observed by confocal microscopy. Scale bar 10 µm. G , Immunoblots and calculated levels (bottom) of HER3, pY845 EGFR, EGFR, pT222 MK2, MK2, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells. Data are expressed as mean of pY845EGFR/EGFR, pT222 MK2/MK2 and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: unpaired t-test for pY845 EGFR; Mann-Whitney test for HER3, EGFR, pT222 MK2, pT180/pY182 p38 MAPK.

    Journal: bioRxiv

    Article Title: Ezrin defines TSC1 activation at endosomal compartments through EGFR-AKT signaling

    doi: 10.1101/2024.05.03.592332

    Figure Lengend Snippet: A, Bubble plot representing the enrichment analysis of 530 DEGs performed in SILAC Phosphoproteomics data. Color and x axis represent minus logarithms of p Value. The size represents numbers of genes enriched in the indicated data. B , Physical interactions, obtained by GeneMANIA, highlight Ezrin and EGFR binding. C , Volcano plot of DEGs, with up-regulated EGFR and down-regulated MAP2 and ERBB2 (no threshold on Log2FC and 0.05 threshold on -Log10FDR). Legend: red dot, up-regulated gene; blue dot, down-regulated gene; grey dot, not significant gene. D, Co-IP data for Ezrin-EGFR interaction. For the co-IP analyses, was used Ezrin antibody, conjugated with beads, and immunoblotted with EGFR antibody for WT and EZR −/− (left) and HeLa EZR T567D and EZR T567A (right) HeLa cells, respectively. Schematic representation of HeLa EZR T567D and EZR T567A co-IP (bottom). E , Confocal microscopy images showing EGFR (green) and EZR (red) co-localization on the membrane in HeLa WT cells (left) and magnified views of the regions are provided (right). Scale bar 10 µm (magnification 1 µm). Representative plots of co-localization profiles on the membrane between EGFR (green) and EZR (red). F , Immunofluorescent labelling images of EGFR in HeLa WT and EZR −/− cells, observed by confocal microscopy. Scale bar 10 µm. G , Immunoblots and calculated levels (bottom) of HER3, pY845 EGFR, EGFR, pT222 MK2, MK2, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells. Data are expressed as mean of pY845EGFR/EGFR, pT222 MK2/MK2 and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: unpaired t-test for pY845 EGFR; Mann-Whitney test for HER3, EGFR, pT222 MK2, pT180/pY182 p38 MAPK.

    Article Snippet: For Western blot analysis, the following antibodies were used: mouse anti-NBR1 (1:1000, Abnova MO1), rabbit anti-LAMP1 (1:500, Sigma L1418), mouse anti-Ezrin (1:1000, Novex 357300), mouse anti-SQSTM1/P62 (1:1000, Abcam ab56416), rabbit anti-Cathepsin D (1:1000, Cell Signaling 2284), rabbit anti-LC3 (1:1000, Novus NB100-2220), mouse anti-GAPDH (1:1000, Santa Cruz SC-32233), rabbit anti-HER2/ErbB2 (1:1000, Cell Signaling 2165), rabbit anti-HER3/ErbB3 (1:1000, Cell Signaling 12708), rabbit anti-phospho-EGF receptor (Tyr845) (1:1000, Cell Signaling 6963), rabbit anti-EGF receptor (1:1000, Cell Signaling 4267), rabbit anti-MAPKAPK-2 (1:1000, Cell Signaling 3042), rabbit anti-phospho-MAPKAPK-2 (Thr222) (1:1000, Cell Signaling 3316), rabbit anti-p38 MAPK (1:1000, Cell Signaling 8690), rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:1000, Cell Signaling 4511), rabbit anti-ZO1 (1:1000, Abcam ab216880), mouse anti-EEA1 (1:1000, BD 610457), rabbit anti-Tuberin/TSC2 (1:1000, Cell Signaling 4308), rabbit anti-phospho-Tuberin/TSC2 (Ser939) (1:1000, Cell Signaling 3615), rabbit anti-phospho-Tuberin/TSC2 (Thr1462) (1:1000, Cell Signaling 3617), rabbit anti-p70 S6 Kinase (1:1000, Cell Signaling 9202), mouse anti-phospho-p70 S6 Kinase (Thr389) (1:1000, Cell Signaling 9206), rabbit anti-Akt (1:1000, Cell Signaling 9272), rabbit anti-phospho-Akt (Ser473) (1:1000, Cell Signaling 4060), rabbit anti-4E-BP1 (1:1000, Cell Signaling 9644), rabbit anti-phospho-4E-BP1 (Ser65) (1:1000, Cell Signaling 9456), rabbit anti-phospho-4E-BP1 (Thr37/46) (1:1000, Cell Signaling 2855), rabbit anti-Hamartin/TSC1 (1:1000, Cell Signaling 6935), mouse anti-EGFR (1:500, Santa Cruz sc-120), mouse anti-p-EGFR (1:500, Santa Cruz sc-57542).

    Techniques: Binding Assay, Co-Immunoprecipitation Assay, Confocal Microscopy, Membrane, Western Blot, MANN-WHITNEY

    A , Live cell imaging and model for EGFR (green) translocation from the membrane to the endosomes in HeLa WT (top) and EZR −/− (bottom) cells without EGF stimulation (T0) and with a progressive EGF stimulation (from T10’’ to T60’’). White boxes are magnifications that depict EGFR protein migration. Scale bar 1 µm). Please refer to Video 1. B , IEM (anti-GFP immunolabelling) of cycloheximide treated HeLa WT, WT + EGF, EZR −/− and EZR −/− + EGF cells expressing EGFR-GFP. Endosomes containing EGFR is shown in green. Scale bar 200 nm. Quantitative analysis (right) of EGFR positive endosomes expressed as mean ± SEM. Statistical test: generalized Linear Model with Likelihood Ratio (Poisson Regression). C , immunoblots and calculated levels (bottom) of HER2, pY845 EGFR, EGFR, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells with (+) and without (-) EGF stimulation. Data are expressed as mean of pY845EGFR/EGFR and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: Unpaired t-test for HER2 WT, HER2 EZR −/− , pY845 EGFR EZR −/− , EGFR WT, EGFR EZR −/− , pT180/pY182 p38 MAPK WT, pT180/pY182 p38 MAPK EZR −/− ; unpaired t-test with Welch’s correction for pY845 EGFR WT. D , Representative immunoblots of EGFR in membrane (top) and endosomes (bottom) proteins in HeLa WT and EZR −/− with (+) and without (-) EGF stimulation. ZO-1 and EEA1 are used as membrane and endosomes extraction control, respectively. GAPDH are used as loading control.

    Journal: bioRxiv

    Article Title: Ezrin defines TSC1 activation at endosomal compartments through EGFR-AKT signaling

    doi: 10.1101/2024.05.03.592332

    Figure Lengend Snippet: A , Live cell imaging and model for EGFR (green) translocation from the membrane to the endosomes in HeLa WT (top) and EZR −/− (bottom) cells without EGF stimulation (T0) and with a progressive EGF stimulation (from T10’’ to T60’’). White boxes are magnifications that depict EGFR protein migration. Scale bar 1 µm). Please refer to Video 1. B , IEM (anti-GFP immunolabelling) of cycloheximide treated HeLa WT, WT + EGF, EZR −/− and EZR −/− + EGF cells expressing EGFR-GFP. Endosomes containing EGFR is shown in green. Scale bar 200 nm. Quantitative analysis (right) of EGFR positive endosomes expressed as mean ± SEM. Statistical test: generalized Linear Model with Likelihood Ratio (Poisson Regression). C , immunoblots and calculated levels (bottom) of HER2, pY845 EGFR, EGFR, pT180/pY182 p38 MAPK and P38 MAPK in HeLa WT and EZR −/− cells with (+) and without (-) EGF stimulation. Data are expressed as mean of pY845EGFR/EGFR and pT180/pY182 p38 MAPK/P38 MAPK ratio ± SEM (n=3 experiments at least). GAPDH was used as loading control. Statistical test: Unpaired t-test for HER2 WT, HER2 EZR −/− , pY845 EGFR EZR −/− , EGFR WT, EGFR EZR −/− , pT180/pY182 p38 MAPK WT, pT180/pY182 p38 MAPK EZR −/− ; unpaired t-test with Welch’s correction for pY845 EGFR WT. D , Representative immunoblots of EGFR in membrane (top) and endosomes (bottom) proteins in HeLa WT and EZR −/− with (+) and without (-) EGF stimulation. ZO-1 and EEA1 are used as membrane and endosomes extraction control, respectively. GAPDH are used as loading control.

    Article Snippet: For Western blot analysis, the following antibodies were used: mouse anti-NBR1 (1:1000, Abnova MO1), rabbit anti-LAMP1 (1:500, Sigma L1418), mouse anti-Ezrin (1:1000, Novex 357300), mouse anti-SQSTM1/P62 (1:1000, Abcam ab56416), rabbit anti-Cathepsin D (1:1000, Cell Signaling 2284), rabbit anti-LC3 (1:1000, Novus NB100-2220), mouse anti-GAPDH (1:1000, Santa Cruz SC-32233), rabbit anti-HER2/ErbB2 (1:1000, Cell Signaling 2165), rabbit anti-HER3/ErbB3 (1:1000, Cell Signaling 12708), rabbit anti-phospho-EGF receptor (Tyr845) (1:1000, Cell Signaling 6963), rabbit anti-EGF receptor (1:1000, Cell Signaling 4267), rabbit anti-MAPKAPK-2 (1:1000, Cell Signaling 3042), rabbit anti-phospho-MAPKAPK-2 (Thr222) (1:1000, Cell Signaling 3316), rabbit anti-p38 MAPK (1:1000, Cell Signaling 8690), rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:1000, Cell Signaling 4511), rabbit anti-ZO1 (1:1000, Abcam ab216880), mouse anti-EEA1 (1:1000, BD 610457), rabbit anti-Tuberin/TSC2 (1:1000, Cell Signaling 4308), rabbit anti-phospho-Tuberin/TSC2 (Ser939) (1:1000, Cell Signaling 3615), rabbit anti-phospho-Tuberin/TSC2 (Thr1462) (1:1000, Cell Signaling 3617), rabbit anti-p70 S6 Kinase (1:1000, Cell Signaling 9202), mouse anti-phospho-p70 S6 Kinase (Thr389) (1:1000, Cell Signaling 9206), rabbit anti-Akt (1:1000, Cell Signaling 9272), rabbit anti-phospho-Akt (Ser473) (1:1000, Cell Signaling 4060), rabbit anti-4E-BP1 (1:1000, Cell Signaling 9644), rabbit anti-phospho-4E-BP1 (Ser65) (1:1000, Cell Signaling 9456), rabbit anti-phospho-4E-BP1 (Thr37/46) (1:1000, Cell Signaling 2855), rabbit anti-Hamartin/TSC1 (1:1000, Cell Signaling 6935), mouse anti-EGFR (1:500, Santa Cruz sc-120), mouse anti-p-EGFR (1:500, Santa Cruz sc-57542).

    Techniques: Live Cell Imaging, Translocation Assay, Membrane, Migration, Expressing, Western Blot, Extraction

    Treatment of cells with Aβ led to increased phospho/total p38 MAPK ratio, while blocking p38 MAPK activity with SB203580 decreased p53 activity in A549 cells and increased ACh levels in the media of both cell lines. Cells (0.2 × 10) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight. The cells were then treated as indicated for 72 h with Aβ 1–40/42 or fragments (10 μM) ± SB203580 (SB, 20 μM), and then, the p38 MAPK assay ( A ) and Western blotting ( B ) were carried out (Materials and Methods). The p53 activity was measured in A549 cells ( C ). Cells transfected with either control or p53 siRNA were treated for 72 h with Aβ 1–40/42 or fragments ± SB203580; then, the levels of ACh ( D – F ) were measured as described in the Materials and Methods Section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to the control of each cell line ( A ) or to control in the absence of SB203580 ( C – F ) using the GraphPad 10.1.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks indicate a statistically significant difference from control. Absence of asterisks indicates no significance, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple comparison test, * p < 0.05, ** p < 0.01.

    Journal: International Journal of Molecular Sciences

    Article Title: Amyloid Beta Leads to Decreased Acetylcholine Levels and Non-Small Cell Lung Cancer Cell Survival via a Mechanism That Involves p38 Mitogen-Activated Protein Kinase and Protein Kinase C in a p53-Dependent and -Independent Manner

    doi: 10.3390/ijms25095033

    Figure Lengend Snippet: Treatment of cells with Aβ led to increased phospho/total p38 MAPK ratio, while blocking p38 MAPK activity with SB203580 decreased p53 activity in A549 cells and increased ACh levels in the media of both cell lines. Cells (0.2 × 10) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight. The cells were then treated as indicated for 72 h with Aβ 1–40/42 or fragments (10 μM) ± SB203580 (SB, 20 μM), and then, the p38 MAPK assay ( A ) and Western blotting ( B ) were carried out (Materials and Methods). The p53 activity was measured in A549 cells ( C ). Cells transfected with either control or p53 siRNA were treated for 72 h with Aβ 1–40/42 or fragments ± SB203580; then, the levels of ACh ( D – F ) were measured as described in the Materials and Methods Section. Data from five independent assays, each carried out in triplicate, were averaged, normalized, and expressed as fold change relative to the control of each cell line ( A ) or to control in the absence of SB203580 ( C – F ) using the GraphPad 10.1.1 software. The graphs summarize the results expressed as means ± SD (n = 5). Asterisks indicate a statistically significant difference from control. Absence of asterisks indicates no significance, Mann–Whitney test. Statistical differences between different groups were analyzed by an ordinary one-way analysis of variance (ANOVA) followed by Tukey’s post hoc multiple comparison test, * p < 0.05, ** p < 0.01.

    Article Snippet: SignalSilence p53 siRNA I (6231), SignalSilence control siRNA (Unconjugated, 6568), rabbit p53 antibody (9282), p38 MAPK antibody (9212) that detects endogenous levels of total p38α/β/γ MAPK, phospho-p38 MAPK (Thr180/Tyr182) antibody (9211), PKCα antibody (2056), and SB203580 (5633S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Blocking Assay, Activity Assay, Western Blot, Transfection, Software, MANN-WHITNEY, Comparison

    Incubation of cells with ACh reduced the activity of p53 in A549 cells, diminished the activity of p38 MAPK, and enhanced the PKC activity in both A549 and H1299 cells, while co-incubation with Aβ reduced those effects. Cells (0.2 × 10 5 ) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight (Control, 0 hour). The cells were then treated as indicated with ACh (100 nM) and in combination with Aβ 1–40/42 or fragments (10 μM). The p53 activity ( A ) in A549 cells and the activity of p38 MAPK ( B , C ) and PKC ( D , E ) were measured as described in the Materials and Methods Section. The data were expressed as the percentage of control by expressing each point relative to the control (set to 100%). The data were then plotted as a function of time using the GraphPad Prism 10.1.1 software. Data were expressed as the mean ± S.D. of three independent experiments, each carried out in triplicate.

    Journal: International Journal of Molecular Sciences

    Article Title: Amyloid Beta Leads to Decreased Acetylcholine Levels and Non-Small Cell Lung Cancer Cell Survival via a Mechanism That Involves p38 Mitogen-Activated Protein Kinase and Protein Kinase C in a p53-Dependent and -Independent Manner

    doi: 10.3390/ijms25095033

    Figure Lengend Snippet: Incubation of cells with ACh reduced the activity of p53 in A549 cells, diminished the activity of p38 MAPK, and enhanced the PKC activity in both A549 and H1299 cells, while co-incubation with Aβ reduced those effects. Cells (0.2 × 10 5 ) were grown in 10% FBS-supplemented media for 24 h and then serum-starved overnight (Control, 0 hour). The cells were then treated as indicated with ACh (100 nM) and in combination with Aβ 1–40/42 or fragments (10 μM). The p53 activity ( A ) in A549 cells and the activity of p38 MAPK ( B , C ) and PKC ( D , E ) were measured as described in the Materials and Methods Section. The data were expressed as the percentage of control by expressing each point relative to the control (set to 100%). The data were then plotted as a function of time using the GraphPad Prism 10.1.1 software. Data were expressed as the mean ± S.D. of three independent experiments, each carried out in triplicate.

    Article Snippet: SignalSilence p53 siRNA I (6231), SignalSilence control siRNA (Unconjugated, 6568), rabbit p53 antibody (9282), p38 MAPK antibody (9212) that detects endogenous levels of total p38α/β/γ MAPK, phospho-p38 MAPK (Thr180/Tyr182) antibody (9211), PKCα antibody (2056), and SB203580 (5633S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Incubation, Activity Assay, Expressing, Software

    Representation of the main hypothesis and findings of this study. Aβ decreases the levels of ACh in the media via the activation of p53/AChE and p38 MAPK and/or via blocking the activity of PKC, leading to decreased cell survival. In this model, ACh reverses the effects of Aβ.

    Journal: International Journal of Molecular Sciences

    Article Title: Amyloid Beta Leads to Decreased Acetylcholine Levels and Non-Small Cell Lung Cancer Cell Survival via a Mechanism That Involves p38 Mitogen-Activated Protein Kinase and Protein Kinase C in a p53-Dependent and -Independent Manner

    doi: 10.3390/ijms25095033

    Figure Lengend Snippet: Representation of the main hypothesis and findings of this study. Aβ decreases the levels of ACh in the media via the activation of p53/AChE and p38 MAPK and/or via blocking the activity of PKC, leading to decreased cell survival. In this model, ACh reverses the effects of Aβ.

    Article Snippet: SignalSilence p53 siRNA I (6231), SignalSilence control siRNA (Unconjugated, 6568), rabbit p53 antibody (9282), p38 MAPK antibody (9212) that detects endogenous levels of total p38α/β/γ MAPK, phospho-p38 MAPK (Thr180/Tyr182) antibody (9211), PKCα antibody (2056), and SB203580 (5633S) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Activation Assay, Blocking Assay, Activity Assay

    Western blotting analysis of the relative expression of MAPK signaling pathway components in the retina in the different intervention groups. The rats were subjected to IOP elevation, and then received the intravitreal injection of DMSO, the CK2 inhibitor TBB, the CK2 inhibitor DMAT, and the macrophage activator ZYM. Rats that were not subjected to IOP elevation and were intravitreally injected with DMSO served as intact controls. (A) The relative protein expression levels of p38 and p-p38 were examined by western blotting. (B) The relative protein expression levels of Erk p44/p42 and p-p44/p42 were detected by western blotting. The results are presented as histograms after normalization to β-actin or GAPDH. The data are presented as mean ± SD ( n = 5). * P < 0.05 (one-way analysis of variance among multiple groups followed by post hoc Bonferroni test). CK2: Casein kinase-2; DMAT: 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole; DMSO: dimethylsulfoxide; Erk: extracellular-signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IOP: intraocular pressure; MAPK: mitogen-activated protein kinase; RGCs: retinal ganglion cells; TBB: 4,5,6,7-tetrabromo-2-azabenzimidazole; ZYM: zymosan.

    Journal: Neural Regeneration Research

    Article Title: Casein kinase-2 inhibition promotes retinal ganglion cell survival after acute intraocular pressure elevation

    doi: 10.4103/1673-5374.385310

    Figure Lengend Snippet: Western blotting analysis of the relative expression of MAPK signaling pathway components in the retina in the different intervention groups. The rats were subjected to IOP elevation, and then received the intravitreal injection of DMSO, the CK2 inhibitor TBB, the CK2 inhibitor DMAT, and the macrophage activator ZYM. Rats that were not subjected to IOP elevation and were intravitreally injected with DMSO served as intact controls. (A) The relative protein expression levels of p38 and p-p38 were examined by western blotting. (B) The relative protein expression levels of Erk p44/p42 and p-p44/p42 were detected by western blotting. The results are presented as histograms after normalization to β-actin or GAPDH. The data are presented as mean ± SD ( n = 5). * P < 0.05 (one-way analysis of variance among multiple groups followed by post hoc Bonferroni test). CK2: Casein kinase-2; DMAT: 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole; DMSO: dimethylsulfoxide; Erk: extracellular-signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IOP: intraocular pressure; MAPK: mitogen-activated protein kinase; RGCs: retinal ganglion cells; TBB: 4,5,6,7-tetrabromo-2-azabenzimidazole; ZYM: zymosan.

    Article Snippet: Next, 50 μg of total protein was resolved by SDS-polyacrylamide gel electrophoresis (Bio-Rad, Hercules, CA, USA), transferred onto a nitrocellulose membrane (Amersham Biosciences, Little Chalfont, UK), blocked with 5% skim milk in 0.05% Tween 20 (ICN, Erie, PA, USA) in Tris-buffered saline, and probed with rabbit anti-extracellular-signal-regulated kinase (Erk) p44/p42 antibody (1:1000, Cell Signaling Technology, Beverly, MA, USA, Cat# 4695, RRID: AB_390779), rabbit anti-phospho-Erk p44/p42 antibody (1:1000, Cell Signaling Technology, Cat# 8544, RRID: AB_11127856), rabbit anti-p38 antibody (1:1000, Cell Signaling Technology, Cat# 14451, RRID: AB_2798482), or rabbit anti-phospho-p38 antibody (1:1000, Cell Signaling Technology, Cat# 4511, RRID: AB_2139682) in 3% bovine serum albumin at 4°C overnight.

    Techniques: Western Blot, Expressing, Injection

    LPA 1 antagonist attenuates NF-κB upregulation and MAPK activation but enhances PI3K/Akt activation after pMCAO challenge. Mice were challenged with pMCAO. AM152 (5 mg/kg, p.o. ) was then administered at one hour after occlusion. (A) Effects of AM152 on NF-κB expression in activated microglia were determined by NF-κB(p65)/Iba1 double immunofluorescence at 1 day after pMCAO challenge. Representative images of NF-κB(p65) /Iba1-double immunopositive cells in ischemic core regions and quantification are shown. Scale bar, 50 μm. n=4 mice per group. *** p <0.001 versus sham. ### p <0.001 versus vehicle-administered pMCAO group. (B) Effects of AM152 on phosphorylation of MAPKs (ERK1/2, p38, and JNK) and PI3K/Akt were determined by Western blot analysis at 1 day after pMCAO challenge. Representative Western blots and quantification are shown. n=3 mice per group. * p <0.05, ** p <0.01, and *** p <0.001 versus sham. # p <0.05, ## p <0.01 and ### p <0.001 versus vehicle-administered pMCAO group.

    Journal: Biomolecules & Therapeutics

    Article Title: Lysophosphatidic Acid Receptor 1 Plays a Pathogenic Role in Permanent Brain Ischemic Stroke by Modulating Neuroinflammatory Responses

    doi: 10.4062/biomolther.2024.052

    Figure Lengend Snippet: LPA 1 antagonist attenuates NF-κB upregulation and MAPK activation but enhances PI3K/Akt activation after pMCAO challenge. Mice were challenged with pMCAO. AM152 (5 mg/kg, p.o. ) was then administered at one hour after occlusion. (A) Effects of AM152 on NF-κB expression in activated microglia were determined by NF-κB(p65)/Iba1 double immunofluorescence at 1 day after pMCAO challenge. Representative images of NF-κB(p65) /Iba1-double immunopositive cells in ischemic core regions and quantification are shown. Scale bar, 50 μm. n=4 mice per group. *** p <0.001 versus sham. ### p <0.001 versus vehicle-administered pMCAO group. (B) Effects of AM152 on phosphorylation of MAPKs (ERK1/2, p38, and JNK) and PI3K/Akt were determined by Western blot analysis at 1 day after pMCAO challenge. Representative Western blots and quantification are shown. n=3 mice per group. * p <0.05, ** p <0.01, and *** p <0.001 versus sham. # p <0.05, ## p <0.01 and ### p <0.001 versus vehicle-administered pMCAO group.

    Article Snippet: These membranes were incubated with primary antibodies against cleaved caspase-3, Bcl-2, phospho-ERK1/2, ERK1/2, phospho-JNK, JNK, phospho-p38, p38, phospho-Akt, Akt (1:1,000, Cell signaling Technology, Danvers, MA, USA), and β-actin (1:5,000, Sigma-Aldrich) overnight at 4°C, followed by incubation with respective secondary antibodies (1:10,000, Jackson ImmunoResearch) for 2 h at R/T.

    Techniques: Activation Assay, Expressing, Immunofluorescence, Western Blot