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


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    Cell Signaling Technology Inc rabbit polyclonal anti phospho p44 42 mapk erk1 2
    Rabbit Polyclonal Anti Phospho P44 42 Mapk Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti phospho p44 42 mapk erk1 2/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    phospho camkii  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho camkii
    JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05
    Phospho Camkii, 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 camkii/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    phospho camkii - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "JOSD2 mediates isoprenaline-induced heart failure by deubiquitinating CaMKIIδ in cardiomyocytes"

    Article Title: JOSD2 mediates isoprenaline-induced heart failure by deubiquitinating CaMKIIδ in cardiomyocytes

    Journal: Cellular and Molecular Life Sciences: CMLS

    doi: 10.1007/s00018-023-05037-7

    JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05
    Figure Legend Snippet: JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05

    Techniques Used: Activation Assay, Infection, Plasmid Preparation, Injection, Staining, Western Blot

    JOSD2 interacts CaMKIIδ directly and promotes CaMKIIδ activation. A-B MS/MS spectrum of the peptide showing FTDEYQLFEELGK and ESTESSNTTIEDEDVK from CaMKIIδ. Single-letter abbreviations: K, Lys; V, Val; I, Ile; G, Gly; Q, Gln; L, Leu; N, Asn; E, Glu; T, Thr; S, Ser; F, Phe; D, Asp; Y, Tyr. m/z, mass/charge ratio. C The complex of JOSD2 and CaMKIIδ in NRVMs were detected by Coimmunoprecipitation. NRVMs were transfected with Flag-tagged JOSD2. Lysates from cells were immunoprecipitated with an anti-Flag antibody. CaMKIIδ levels were detected by immunoblotting. IgG was used as a control for IP. D In HEK-293 T cells which express a few endogenous CaMKII, Flag-tagged JOSD2 and GFP-tagged CaMKIIδ were transfected. Lysates from cells were immunoprecipitated with anti-Flag antibody. GFP levels were detected by immunoblotting. IgG was used as a control for IP. E Representative western blot for CaMKIIδ and p-CaMKIIδ in NRVMs with gradient levels of JOSD2. F–G JOSD2 was silenced or overexpressed in NRVMs. Representative western blotting for CaMKIIδ and p-CaMKIIδ in NRVMs. GAPDH was used as a loading control. (H-I) Representative western blotting for CaMKIIδ and p-CaMKIIδ in heart tissues from ISO- and MI-induced mice. J–K Confocal images (J)and representative recordings (K) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with si-JOSD2 or si-NC before ISO stimulation for 24 h. n = 3 cells in each group. [Scale bar, 50 μm]. L–M Confocal images (L) and representative recordings (M) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with Flag-JOSD2 or empty vector (EV) before ISO stimulation for 24 h. n = 6 cells in each group. [Scale bar, 50 μm]
    Figure Legend Snippet: JOSD2 interacts CaMKIIδ directly and promotes CaMKIIδ activation. A-B MS/MS spectrum of the peptide showing FTDEYQLFEELGK and ESTESSNTTIEDEDVK from CaMKIIδ. Single-letter abbreviations: K, Lys; V, Val; I, Ile; G, Gly; Q, Gln; L, Leu; N, Asn; E, Glu; T, Thr; S, Ser; F, Phe; D, Asp; Y, Tyr. m/z, mass/charge ratio. C The complex of JOSD2 and CaMKIIδ in NRVMs were detected by Coimmunoprecipitation. NRVMs were transfected with Flag-tagged JOSD2. Lysates from cells were immunoprecipitated with an anti-Flag antibody. CaMKIIδ levels were detected by immunoblotting. IgG was used as a control for IP. D In HEK-293 T cells which express a few endogenous CaMKII, Flag-tagged JOSD2 and GFP-tagged CaMKIIδ were transfected. Lysates from cells were immunoprecipitated with anti-Flag antibody. GFP levels were detected by immunoblotting. IgG was used as a control for IP. E Representative western blot for CaMKIIδ and p-CaMKIIδ in NRVMs with gradient levels of JOSD2. F–G JOSD2 was silenced or overexpressed in NRVMs. Representative western blotting for CaMKIIδ and p-CaMKIIδ in NRVMs. GAPDH was used as a loading control. (H-I) Representative western blotting for CaMKIIδ and p-CaMKIIδ in heart tissues from ISO- and MI-induced mice. J–K Confocal images (J)and representative recordings (K) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with si-JOSD2 or si-NC before ISO stimulation for 24 h. n = 3 cells in each group. [Scale bar, 50 μm]. L–M Confocal images (L) and representative recordings (M) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with Flag-JOSD2 or empty vector (EV) before ISO stimulation for 24 h. n = 6 cells in each group. [Scale bar, 50 μm]

    Techniques Used: Activation Assay, Tandem Mass Spectroscopy, Transfection, Immunoprecipitation, Western Blot, Fluorescence, Plasmid Preparation

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


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc rabbit polyclonal anti phospho p44 42 mapk erk1 2
    Rabbit Polyclonal Anti Phospho P44 42 Mapk Erk1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti phospho p44 42 mapk erk1 2/product/Cell Signaling Technology Inc
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    phospho camkii  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho camkii
    JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05
    Phospho Camkii, 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 camkii/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho camkii - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "JOSD2 mediates isoprenaline-induced heart failure by deubiquitinating CaMKIIδ in cardiomyocytes"

    Article Title: JOSD2 mediates isoprenaline-induced heart failure by deubiquitinating CaMKIIδ in cardiomyocytes

    Journal: Cellular and Molecular Life Sciences: CMLS

    doi: 10.1007/s00018-023-05037-7

    JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05
    Figure Legend Snippet: JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05

    Techniques Used: Activation Assay, Infection, Plasmid Preparation, Injection, Staining, Western Blot

    JOSD2 interacts CaMKIIδ directly and promotes CaMKIIδ activation. A-B MS/MS spectrum of the peptide showing FTDEYQLFEELGK and ESTESSNTTIEDEDVK from CaMKIIδ. Single-letter abbreviations: K, Lys; V, Val; I, Ile; G, Gly; Q, Gln; L, Leu; N, Asn; E, Glu; T, Thr; S, Ser; F, Phe; D, Asp; Y, Tyr. m/z, mass/charge ratio. C The complex of JOSD2 and CaMKIIδ in NRVMs were detected by Coimmunoprecipitation. NRVMs were transfected with Flag-tagged JOSD2. Lysates from cells were immunoprecipitated with an anti-Flag antibody. CaMKIIδ levels were detected by immunoblotting. IgG was used as a control for IP. D In HEK-293 T cells which express a few endogenous CaMKII, Flag-tagged JOSD2 and GFP-tagged CaMKIIδ were transfected. Lysates from cells were immunoprecipitated with anti-Flag antibody. GFP levels were detected by immunoblotting. IgG was used as a control for IP. E Representative western blot for CaMKIIδ and p-CaMKIIδ in NRVMs with gradient levels of JOSD2. F–G JOSD2 was silenced or overexpressed in NRVMs. Representative western blotting for CaMKIIδ and p-CaMKIIδ in NRVMs. GAPDH was used as a loading control. (H-I) Representative western blotting for CaMKIIδ and p-CaMKIIδ in heart tissues from ISO- and MI-induced mice. J–K Confocal images (J)and representative recordings (K) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with si-JOSD2 or si-NC before ISO stimulation for 24 h. n = 3 cells in each group. [Scale bar, 50 μm]. L–M Confocal images (L) and representative recordings (M) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with Flag-JOSD2 or empty vector (EV) before ISO stimulation for 24 h. n = 6 cells in each group. [Scale bar, 50 μm]
    Figure Legend Snippet: JOSD2 interacts CaMKIIδ directly and promotes CaMKIIδ activation. A-B MS/MS spectrum of the peptide showing FTDEYQLFEELGK and ESTESSNTTIEDEDVK from CaMKIIδ. Single-letter abbreviations: K, Lys; V, Val; I, Ile; G, Gly; Q, Gln; L, Leu; N, Asn; E, Glu; T, Thr; S, Ser; F, Phe; D, Asp; Y, Tyr. m/z, mass/charge ratio. C The complex of JOSD2 and CaMKIIδ in NRVMs were detected by Coimmunoprecipitation. NRVMs were transfected with Flag-tagged JOSD2. Lysates from cells were immunoprecipitated with an anti-Flag antibody. CaMKIIδ levels were detected by immunoblotting. IgG was used as a control for IP. D In HEK-293 T cells which express a few endogenous CaMKII, Flag-tagged JOSD2 and GFP-tagged CaMKIIδ were transfected. Lysates from cells were immunoprecipitated with anti-Flag antibody. GFP levels were detected by immunoblotting. IgG was used as a control for IP. E Representative western blot for CaMKIIδ and p-CaMKIIδ in NRVMs with gradient levels of JOSD2. F–G JOSD2 was silenced or overexpressed in NRVMs. Representative western blotting for CaMKIIδ and p-CaMKIIδ in NRVMs. GAPDH was used as a loading control. (H-I) Representative western blotting for CaMKIIδ and p-CaMKIIδ in heart tissues from ISO- and MI-induced mice. J–K Confocal images (J)and representative recordings (K) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with si-JOSD2 or si-NC before ISO stimulation for 24 h. n = 3 cells in each group. [Scale bar, 50 μm]. L–M Confocal images (L) and representative recordings (M) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with Flag-JOSD2 or empty vector (EV) before ISO stimulation for 24 h. n = 6 cells in each group. [Scale bar, 50 μm]

    Techniques Used: Activation Assay, Tandem Mass Spectroscopy, Transfection, Immunoprecipitation, Western Blot, Fluorescence, Plasmid Preparation

    phospho p70s6kinase ser371  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc phospho p70s6kinase ser371
    Phospho P70s6kinase Ser371, 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 p70s6kinase ser371/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    phospho p38 mapk thr180 tyr182 rabbit mab  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 rabbit mab
    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Phospho P38 Mapk Thr180 Tyr182 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho p38 mapk thr180 tyr182 rabbit mab/product/Cell Signaling Technology Inc
    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, 2024-06
    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)


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

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

    Journal: Neural Regeneration Research

    doi: 10.4103/1673-5374.391193

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

    Techniques Used: In Vivo, In Vitro

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

    Techniques Used: Light Microscopy, Comparison

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

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

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

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

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

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

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

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

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

    Techniques Used:

    phospho inositol  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho inositol
    Phospho Inositol, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho inositol  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho inositol
    Phospho Inositol, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho c jun  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho c jun
    Inhibitory effects of ATX on PM 2.5 -induced transcription factor, AP-1, pro-inflammatory cytokines and MMPs. (A and B) Western blot assay was performed for the detection of (A) protein levels of <t>phospho-c-Jun,</t> c-Jun, c-Fos, and (B) protein levels of IL-1β, MMP-2 and MMP-9. * P<0.05 vs. ATX or PM 2.5 -untreated cells; # P<0.05 vs. PM 2.5 -treated cells. MMPs, matrix metalloproteinases; c-Jun, jun proto-oncogene, AP-1 transcription factor subunit; c-Fos, fos proto-oncogene, AP-1 transcription factor subunit.
    Phospho C Jun, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Protective effects of astaxanthin on particulate matter 2.5‑induced senescence in HaCaT keratinocytes via maintenance of redox homeostasis"

    Article Title: Protective effects of astaxanthin on particulate matter 2.5‑induced senescence in HaCaT keratinocytes via maintenance of redox homeostasis

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2024.12563

    Inhibitory effects of ATX on PM 2.5 -induced transcription factor, AP-1, pro-inflammatory cytokines and MMPs. (A and B) Western blot assay was performed for the detection of (A) protein levels of phospho-c-Jun, c-Jun, c-Fos, and (B) protein levels of IL-1β, MMP-2 and MMP-9. * P<0.05 vs. ATX or PM 2.5 -untreated cells; # P<0.05 vs. PM 2.5 -treated cells. MMPs, matrix metalloproteinases; c-Jun, jun proto-oncogene, AP-1 transcription factor subunit; c-Fos, fos proto-oncogene, AP-1 transcription factor subunit.
    Figure Legend Snippet: Inhibitory effects of ATX on PM 2.5 -induced transcription factor, AP-1, pro-inflammatory cytokines and MMPs. (A and B) Western blot assay was performed for the detection of (A) protein levels of phospho-c-Jun, c-Jun, c-Fos, and (B) protein levels of IL-1β, MMP-2 and MMP-9. * P<0.05 vs. ATX or PM 2.5 -untreated cells; # P<0.05 vs. PM 2.5 -treated cells. MMPs, matrix metalloproteinases; c-Jun, jun proto-oncogene, AP-1 transcription factor subunit; c-Fos, fos proto-oncogene, AP-1 transcription factor subunit.

    Techniques Used: Western Blot

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    JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05
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    Cell Signaling Technology Inc phospho p38 mapk thr180 tyr182 rabbit mab
    Schematic of the experimental timelines of in vivo (top) and in vitro (bottom) modeling. MAPK: Mitogen-activated protein kinase; SB202190: <t>p38</t> <t>MAPK</t> inhibitor.
    Phospho P38 Mapk Thr180 Tyr182 Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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
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    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.
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    Inhibitory effects of ATX on PM 2.5 -induced transcription factor, AP-1, pro-inflammatory cytokines and MMPs. (A and B) Western blot assay was performed for the detection of (A) protein levels of <t>phospho-c-Jun,</t> c-Jun, c-Fos, and (B) protein levels of IL-1β, MMP-2 and MMP-9. * P<0.05 vs. ATX or PM 2.5 -untreated cells; # P<0.05 vs. PM 2.5 -treated cells. MMPs, matrix metalloproteinases; c-Jun, jun proto-oncogene, AP-1 transcription factor subunit; c-Fos, fos proto-oncogene, AP-1 transcription factor subunit.
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    Image Search Results


    JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: JOSD2 mediates isoprenaline-induced heart failure by deubiquitinating CaMKIIδ in cardiomyocytes

    doi: 10.1007/s00018-023-05037-7

    Figure Lengend Snippet: JOSD2 aggravates ISO-induced myocardial injury via activation of CaMKIIδ. WT mice were infected with AAV9 carrying JOSD2 cDNA (AAV9-JOSD2) or an empty vector (AAV9-NC) through tail vein. Four weeks post-injection, mice were infused with ISO. During the ISO-infused period, KN-93 was administered at 5 mg/kg/day. A Representative M-mode echocardiographic images from mice in each group. B Representative images from harvested heart tissues (scale bar: 2.5 mm). C–I Representative images and quantification of heart tissues stained with WGA (C–D), H&E (E), Sirius Red (F–G) and Masson (H–I) in sections of hearts (scale bar: 100 μm). J The content of ANP in plasma. K mRNA levels of hypertrophy-associated genes and fibrosis-associated genes in mouse heart tissues. Data was normalized to Actb. L Western blot analysis of hypertrophy-associated β-MyHC, ANP and fibrosis-associated COL-1, TGF-β1 in heart tissues. GAPDH was used as the loading control. M Western blot analysis of p-CaMKIIδ, CaMKIIδ in heart tissues. GAPDH was used as the loading control. All quantitative data is presented as Mean ± SEM; n = 6; ns = not significant; * = p < 0.05

    Article Snippet: Phospho-CaMKII (Cat# 12716 s), horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Cat# 7076), and goat anti-rabbit IgG (Cat# 7074) were purchased from Cell Signaling Technology (CST, Danvers, MA, USA).

    Techniques: Activation Assay, Infection, Plasmid Preparation, Injection, Staining, Western Blot

    JOSD2 interacts CaMKIIδ directly and promotes CaMKIIδ activation. A-B MS/MS spectrum of the peptide showing FTDEYQLFEELGK and ESTESSNTTIEDEDVK from CaMKIIδ. Single-letter abbreviations: K, Lys; V, Val; I, Ile; G, Gly; Q, Gln; L, Leu; N, Asn; E, Glu; T, Thr; S, Ser; F, Phe; D, Asp; Y, Tyr. m/z, mass/charge ratio. C The complex of JOSD2 and CaMKIIδ in NRVMs were detected by Coimmunoprecipitation. NRVMs were transfected with Flag-tagged JOSD2. Lysates from cells were immunoprecipitated with an anti-Flag antibody. CaMKIIδ levels were detected by immunoblotting. IgG was used as a control for IP. D In HEK-293 T cells which express a few endogenous CaMKII, Flag-tagged JOSD2 and GFP-tagged CaMKIIδ were transfected. Lysates from cells were immunoprecipitated with anti-Flag antibody. GFP levels were detected by immunoblotting. IgG was used as a control for IP. E Representative western blot for CaMKIIδ and p-CaMKIIδ in NRVMs with gradient levels of JOSD2. F–G JOSD2 was silenced or overexpressed in NRVMs. Representative western blotting for CaMKIIδ and p-CaMKIIδ in NRVMs. GAPDH was used as a loading control. (H-I) Representative western blotting for CaMKIIδ and p-CaMKIIδ in heart tissues from ISO- and MI-induced mice. J–K Confocal images (J)and representative recordings (K) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with si-JOSD2 or si-NC before ISO stimulation for 24 h. n = 3 cells in each group. [Scale bar, 50 μm]. L–M Confocal images (L) and representative recordings (M) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with Flag-JOSD2 or empty vector (EV) before ISO stimulation for 24 h. n = 6 cells in each group. [Scale bar, 50 μm]

    Journal: Cellular and Molecular Life Sciences: CMLS

    Article Title: JOSD2 mediates isoprenaline-induced heart failure by deubiquitinating CaMKIIδ in cardiomyocytes

    doi: 10.1007/s00018-023-05037-7

    Figure Lengend Snippet: JOSD2 interacts CaMKIIδ directly and promotes CaMKIIδ activation. A-B MS/MS spectrum of the peptide showing FTDEYQLFEELGK and ESTESSNTTIEDEDVK from CaMKIIδ. Single-letter abbreviations: K, Lys; V, Val; I, Ile; G, Gly; Q, Gln; L, Leu; N, Asn; E, Glu; T, Thr; S, Ser; F, Phe; D, Asp; Y, Tyr. m/z, mass/charge ratio. C The complex of JOSD2 and CaMKIIδ in NRVMs were detected by Coimmunoprecipitation. NRVMs were transfected with Flag-tagged JOSD2. Lysates from cells were immunoprecipitated with an anti-Flag antibody. CaMKIIδ levels were detected by immunoblotting. IgG was used as a control for IP. D In HEK-293 T cells which express a few endogenous CaMKII, Flag-tagged JOSD2 and GFP-tagged CaMKIIδ were transfected. Lysates from cells were immunoprecipitated with anti-Flag antibody. GFP levels were detected by immunoblotting. IgG was used as a control for IP. E Representative western blot for CaMKIIδ and p-CaMKIIδ in NRVMs with gradient levels of JOSD2. F–G JOSD2 was silenced or overexpressed in NRVMs. Representative western blotting for CaMKIIδ and p-CaMKIIδ in NRVMs. GAPDH was used as a loading control. (H-I) Representative western blotting for CaMKIIδ and p-CaMKIIδ in heart tissues from ISO- and MI-induced mice. J–K Confocal images (J)and representative recordings (K) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with si-JOSD2 or si-NC before ISO stimulation for 24 h. n = 3 cells in each group. [Scale bar, 50 μm]. L–M Confocal images (L) and representative recordings (M) of SR Ca2+ determined by the Fluo-4 AM fluorescence signal in cardiomyocytes transfected with Flag-JOSD2 or empty vector (EV) before ISO stimulation for 24 h. n = 6 cells in each group. [Scale bar, 50 μm]

    Article Snippet: Phospho-CaMKII (Cat# 12716 s), horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Cat# 7076), and goat anti-rabbit IgG (Cat# 7074) were purchased from Cell Signaling Technology (CST, Danvers, MA, USA).

    Techniques: Activation Assay, Tandem Mass Spectroscopy, Transfection, Immunoprecipitation, Western Blot, Fluorescence, Plasmid Preparation

    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:

    Inhibitory effects of ATX on PM 2.5 -induced transcription factor, AP-1, pro-inflammatory cytokines and MMPs. (A and B) Western blot assay was performed for the detection of (A) protein levels of phospho-c-Jun, c-Jun, c-Fos, and (B) protein levels of IL-1β, MMP-2 and MMP-9. * P<0.05 vs. ATX or PM 2.5 -untreated cells; # P<0.05 vs. PM 2.5 -treated cells. MMPs, matrix metalloproteinases; c-Jun, jun proto-oncogene, AP-1 transcription factor subunit; c-Fos, fos proto-oncogene, AP-1 transcription factor subunit.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Protective effects of astaxanthin on particulate matter 2.5‑induced senescence in HaCaT keratinocytes via maintenance of redox homeostasis

    doi: 10.3892/etm.2024.12563

    Figure Lengend Snippet: Inhibitory effects of ATX on PM 2.5 -induced transcription factor, AP-1, pro-inflammatory cytokines and MMPs. (A and B) Western blot assay was performed for the detection of (A) protein levels of phospho-c-Jun, c-Jun, c-Fos, and (B) protein levels of IL-1β, MMP-2 and MMP-9. * P<0.05 vs. ATX or PM 2.5 -untreated cells; # P<0.05 vs. PM 2.5 -treated cells. MMPs, matrix metalloproteinases; c-Jun, jun proto-oncogene, AP-1 transcription factor subunit; c-Fos, fos proto-oncogene, AP-1 transcription factor subunit.

    Article Snippet: The following primary antibodies were used: NRF2 (cat. no. sc-722), CAT (cat. no. sc-271803), GPX1/2 (cat. no. sc-133160), cyclin dependent kinase inhibitor 2A (p16) (cat. no. sc-1661), HO-1 (cat. no. sc-390991) and actin (cat. no. sc-8432) were purchased from Santa Cruz Biotechnology, Inc. Phospho-H2A histone family member X (H2A.X; cat. no. 2577), H2A.X (cat. no. 2595), c-Fos (cat. no. 2250), jun proto-oncogene, activator protein-1 (AP-1) transcription factor subunit (c-Jun; cat. no. 9165), phospho-c-Jun (cat. no. 91952) were obtained from Cell Signaling Technology, Inc. Phospho-NRF2 (cat. no. ab76026), interleukin (IL)-1β (cat. no. ab315084), MMP-2 (cat. no. ab92536), MMP-9 (cat. no. ab76003) and TATA-binding protein (TBP) (cat. no. ab818) were purchased from Abcam.

    Techniques: Western Blot