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Mitochondrial reactive oxygen species (ROS) and mitochondrial permeability transition pore (mPTP) opening promote ferroptosis in oxygen-glucose deprivation (OGD)-treated neurons. (A) Representative fluorescence images of MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in the control (Con) and OGD groups, with or without Mito-TEMPO (Mito-T). (B) Quantification of fluorescence intensity for the indicated oxidative stress markers in each condition. (C) Representative fluorescence images and quantification of calcein fluorescence in the control and OGD groups with or without cyclosporin A (CSA), using the calcein–CoCl 2 mPTP quenching assay to assess mPTP opening. (D) Representative flow cytometry histograms and quantification <t>of</t> <t>JC-1</t> staining to assess mitochondrial membrane potential (ΔΨm). Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test. * P < 0.05; ** P < 0.01; *** P < 0.001.
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Mitochondrial reactive oxygen species (ROS) and mitochondrial permeability transition pore (mPTP) opening promote ferroptosis in oxygen-glucose deprivation (OGD)-treated neurons. (A) Representative fluorescence images of MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in the control (Con) and OGD groups, with or without Mito-TEMPO (Mito-T). (B) Quantification of fluorescence intensity for the indicated oxidative stress markers in each condition. (C) Representative fluorescence images and quantification of calcein fluorescence in the control and OGD groups with or without cyclosporin A (CSA), using the calcein–CoCl 2 mPTP quenching assay to assess mPTP opening. (D) Representative flow cytometry histograms and quantification <t>of</t> <t>JC-1</t> staining to assess mitochondrial membrane potential (ΔΨm). Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test. * P < 0.05; ** P < 0.01; *** P < 0.001.
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Mitochondrial reactive oxygen species (ROS) and mitochondrial permeability transition pore (mPTP) opening promote ferroptosis in oxygen-glucose deprivation (OGD)-treated neurons. (A) Representative fluorescence images of MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in the control (Con) and OGD groups, with or without Mito-TEMPO (Mito-T). (B) Quantification of fluorescence intensity for the indicated oxidative stress markers in each condition. (C) Representative fluorescence images and quantification of calcein fluorescence in the control and OGD groups with or without cyclosporin A (CSA), using the calcein–CoCl 2 mPTP quenching assay to assess mPTP opening. (D) Representative flow cytometry histograms and quantification <t>of</t> <t>JC-1</t> staining to assess mitochondrial membrane potential (ΔΨm). Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test. * P < 0.05; ** P < 0.01; *** P < 0.001.
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Image Search Results


Mitochondrial reactive oxygen species (ROS) and mitochondrial permeability transition pore (mPTP) opening promote ferroptosis in oxygen-glucose deprivation (OGD)-treated neurons. (A) Representative fluorescence images of MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in the control (Con) and OGD groups, with or without Mito-TEMPO (Mito-T). (B) Quantification of fluorescence intensity for the indicated oxidative stress markers in each condition. (C) Representative fluorescence images and quantification of calcein fluorescence in the control and OGD groups with or without cyclosporin A (CSA), using the calcein–CoCl 2 mPTP quenching assay to assess mPTP opening. (D) Representative flow cytometry histograms and quantification of JC-1 staining to assess mitochondrial membrane potential (ΔΨm). Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Journal: Research

Article Title: Extracellular Vesicle-Mediated Delivery of Mitochondrial Circular RNA MTCO2 Protects against Cerebral Ischemia by Modulating mPTP-Dependent Ferroptosis

doi: 10.34133/research.1232

Figure Lengend Snippet: Mitochondrial reactive oxygen species (ROS) and mitochondrial permeability transition pore (mPTP) opening promote ferroptosis in oxygen-glucose deprivation (OGD)-treated neurons. (A) Representative fluorescence images of MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in the control (Con) and OGD groups, with or without Mito-TEMPO (Mito-T). (B) Quantification of fluorescence intensity for the indicated oxidative stress markers in each condition. (C) Representative fluorescence images and quantification of calcein fluorescence in the control and OGD groups with or without cyclosporin A (CSA), using the calcein–CoCl 2 mPTP quenching assay to assess mPTP opening. (D) Representative flow cytometry histograms and quantification of JC-1 staining to assess mitochondrial membrane potential (ΔΨm). Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Mitochondrial transmembrane potential was assessed using the lipophilic cation probe JC-1 (MedChemExpress, USA), which accumulates in a potential-dependent manner.

Techniques: Permeability, Fluorescence, Control, Flow Cytometry, Staining, Membrane

Mitochondria-targeted circMTCO2 delivery alleviates oxygen-glucose deprivation (OGD)-induced neuronal ferroptosis. (A) Representative fluorescence images of calcein, MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in Neuro2a (N2a) cells under control or OGD conditions, treated with phosphate-buffered saline (PBS), EV Ctrl , RVG-EV RNA , and RVG-EV mt-RNA . (B) Quantification of fluorescence intensities shown in panel (A). (C) Representative flow cytometry histograms and quantification of JC-1 staining to assess mitochondrial membrane potential (ΔΨm). (D) Cell viability assessed by the Cell Counting Kit-8 (CCK-8) assay. Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test and one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Journal: Research

Article Title: Extracellular Vesicle-Mediated Delivery of Mitochondrial Circular RNA MTCO2 Protects against Cerebral Ischemia by Modulating mPTP-Dependent Ferroptosis

doi: 10.34133/research.1232

Figure Lengend Snippet: Mitochondria-targeted circMTCO2 delivery alleviates oxygen-glucose deprivation (OGD)-induced neuronal ferroptosis. (A) Representative fluorescence images of calcein, MitoSOX, MitoPeDPP, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), and C11-BODIPY (oxidized and reduced forms) in Neuro2a (N2a) cells under control or OGD conditions, treated with phosphate-buffered saline (PBS), EV Ctrl , RVG-EV RNA , and RVG-EV mt-RNA . (B) Quantification of fluorescence intensities shown in panel (A). (C) Representative flow cytometry histograms and quantification of JC-1 staining to assess mitochondrial membrane potential (ΔΨm). (D) Cell viability assessed by the Cell Counting Kit-8 (CCK-8) assay. Scale bars, 5 μm. Data are presented as mean ± standard error of the mean (SEM) ( n = 3 to 4 per group). Statistical analysis was performed using an unpaired 2-tailed Student t test and one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: Mitochondrial transmembrane potential was assessed using the lipophilic cation probe JC-1 (MedChemExpress, USA), which accumulates in a potential-dependent manner.

Techniques: Fluorescence, Control, Saline, Flow Cytometry, Staining, Membrane, Cell Counting, CCK-8 Assay