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Dojindo Labs gsh gssg quantification kit
ATM inhibition in combination with IFN-γ alters glutathione metabolism in NSCLC Cells were treated with IFN-γ (1000 ng/mL) and/or KU-55933 (10 μM) for 24 h, and then the intracellular levels of GSH and GSSG were quantified. The results of total glutathione (A), GSH (B), GSSG (C), <t>and</t> <t>GSH/GSSG</t> ratio (D) are shown. * p < 0.05.
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ATM inhibition in combination with IFN-γ alters glutathione metabolism in NSCLC Cells were treated with IFN-γ (1000 ng/mL) and/or KU-55933 (10 μM) for 24 h, and then the intracellular levels of GSH and GSSG were quantified. The results of total glutathione (A), GSH (B), GSSG (C), <t>and</t> <t>GSH/GSSG</t> ratio (D) are shown. * p < 0.05.
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ATM inhibition in combination with IFN-γ alters glutathione metabolism in NSCLC Cells were treated with IFN-γ (1000 ng/mL) and/or KU-55933 (10 μM) for 24 h, and then the intracellular levels of GSH and GSSG were quantified. The results of total glutathione (A), GSH (B), GSSG (C), <t>and</t> <t>GSH/GSSG</t> ratio (D) are shown. * p < 0.05.
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ATM inhibition in combination with IFN-γ alters glutathione metabolism in NSCLC Cells were treated with IFN-γ (1000 ng/mL) and/or KU-55933 (10 μM) for 24 h, and then the intracellular levels of GSH and GSSG were quantified. The results of total glutathione (A), GSH (B), GSSG (C), <t>and</t> <t>GSH/GSSG</t> ratio (D) are shown. * p < 0.05.
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Rg3@PACVs attenuate oxidative stress and mitochondrial dysfunction in hypoxia/reoxygenation (H/R)-injured lung epithelial cells (A) Reactive oxygen species (ROS) levels detected by flow cytometry using DCFH-DA probe. (B) Intracellular total superoxide dismutase (T-SOD) activity. (C) Malondialdehyde (MDA) content. (D) Glutathione/oxidized glutathione <t>(GSH/GSSG)</t> ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and heme oxygenase-1 (HO-1, red). Nuclei counterstained with DAPI (blue). Scale bars, 100 μm. (F) Mitochondrial membrane potential assessed by JC-1 staining. (G) Super-resolution microscopy of mitochondrial ultrastructure (red) in epithelial cells. Lower panel shows magnified views (∗ represents damaged mitochondria, Scale bars, 5 μm). (H–L) Mitochondrial respiration in BEAS-2B cells under different stimulation conditions was assessed by measuring the oxygen consumption rate (OCR) using a Seahorse XF96 analyzer (H). Key parameters including basal respiration (I), maximal respiration (J), proton leak (K), and ATP production (L) were calculated. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 4 biological replicates).
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Rg3@PACVs attenuate oxidative stress and mitochondrial dysfunction in hypoxia/reoxygenation (H/R)-injured lung epithelial cells (A) Reactive oxygen species (ROS) levels detected by flow cytometry using DCFH-DA probe. (B) Intracellular total superoxide dismutase (T-SOD) activity. (C) Malondialdehyde (MDA) content. (D) Glutathione/oxidized glutathione <t>(GSH/GSSG)</t> ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and heme oxygenase-1 (HO-1, red). Nuclei counterstained with DAPI (blue). Scale bars, 100 μm. (F) Mitochondrial membrane potential assessed by JC-1 staining. (G) Super-resolution microscopy of mitochondrial ultrastructure (red) in epithelial cells. Lower panel shows magnified views (∗ represents damaged mitochondria, Scale bars, 5 μm). (H–L) Mitochondrial respiration in BEAS-2B cells under different stimulation conditions was assessed by measuring the oxygen consumption rate (OCR) using a Seahorse XF96 analyzer (H). Key parameters including basal respiration (I), maximal respiration (J), proton leak (K), and ATP production (L) were calculated. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 4 biological replicates).
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Rg3@PACVs attenuate oxidative stress and mitochondrial dysfunction in hypoxia/reoxygenation (H/R)-injured lung epithelial cells (A) Reactive oxygen species (ROS) levels detected by flow cytometry using DCFH-DA probe. (B) Intracellular total superoxide dismutase (T-SOD) activity. (C) Malondialdehyde (MDA) content. (D) Glutathione/oxidized glutathione <t>(GSH/GSSG)</t> ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and heme oxygenase-1 (HO-1, red). Nuclei counterstained with DAPI (blue). Scale bars, 100 μm. (F) Mitochondrial membrane potential assessed by JC-1 staining. (G) Super-resolution microscopy of mitochondrial ultrastructure (red) in epithelial cells. Lower panel shows magnified views (∗ represents damaged mitochondria, Scale bars, 5 μm). (H–L) Mitochondrial respiration in BEAS-2B cells under different stimulation conditions was assessed by measuring the oxygen consumption rate (OCR) using a Seahorse XF96 analyzer (H). Key parameters including basal respiration (I), maximal respiration (J), proton leak (K), and ATP production (L) were calculated. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 4 biological replicates).
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Rg3@PACVs attenuate oxidative stress and mitochondrial dysfunction in hypoxia/reoxygenation (H/R)-injured lung epithelial cells (A) Reactive oxygen species (ROS) levels detected by flow cytometry using DCFH-DA probe. (B) Intracellular total superoxide dismutase (T-SOD) activity. (C) Malondialdehyde (MDA) content. (D) Glutathione/oxidized glutathione <t>(GSH/GSSG)</t> ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and heme oxygenase-1 (HO-1, red). Nuclei counterstained with DAPI (blue). Scale bars, 100 μm. (F) Mitochondrial membrane potential assessed by JC-1 staining. (G) Super-resolution microscopy of mitochondrial ultrastructure (red) in epithelial cells. Lower panel shows magnified views (∗ represents damaged mitochondria, Scale bars, 5 μm). (H–L) Mitochondrial respiration in BEAS-2B cells under different stimulation conditions was assessed by measuring the oxygen consumption rate (OCR) using a Seahorse XF96 analyzer (H). Key parameters including basal respiration (I), maximal respiration (J), proton leak (K), and ATP production (L) were calculated. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 4 biological replicates).
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Rg3@PACVs attenuate oxidative stress and mitochondrial dysfunction in hypoxia/reoxygenation (H/R)-injured lung epithelial cells (A) Reactive oxygen species (ROS) levels detected by flow cytometry using DCFH-DA probe. (B) Intracellular total superoxide dismutase (T-SOD) activity. (C) Malondialdehyde (MDA) content. (D) Glutathione/oxidized glutathione <t>(GSH/GSSG)</t> ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and heme oxygenase-1 (HO-1, red). Nuclei counterstained with DAPI (blue). Scale bars, 100 μm. (F) Mitochondrial membrane potential assessed by JC-1 staining. (G) Super-resolution microscopy of mitochondrial ultrastructure (red) in epithelial cells. Lower panel shows magnified views (∗ represents damaged mitochondria, Scale bars, 5 μm). (H–L) Mitochondrial respiration in BEAS-2B cells under different stimulation conditions was assessed by measuring the oxygen consumption rate (OCR) using a Seahorse XF96 analyzer (H). Key parameters including basal respiration (I), maximal respiration (J), proton leak (K), and ATP production (L) were calculated. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 4 biological replicates).
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ATM inhibition in combination with IFN-γ alters glutathione metabolism in NSCLC Cells were treated with IFN-γ (1000 ng/mL) and/or KU-55933 (10 μM) for 24 h, and then the intracellular levels of GSH and GSSG were quantified. The results of total glutathione (A), GSH (B), GSSG (C), and GSH/GSSG ratio (D) are shown. * p < 0.05.

Journal: Biochemistry and Biophysics Reports

Article Title: ATM inhibition restores IFN-γ sensitivity and induces ferroptosis in NSCLC via DNA damage response

doi: 10.1016/j.bbrep.2026.102568

Figure Lengend Snippet: ATM inhibition in combination with IFN-γ alters glutathione metabolism in NSCLC Cells were treated with IFN-γ (1000 ng/mL) and/or KU-55933 (10 μM) for 24 h, and then the intracellular levels of GSH and GSSG were quantified. The results of total glutathione (A), GSH (B), GSSG (C), and GSH/GSSG ratio (D) are shown. * p < 0.05.

Article Snippet: Intracellular glutathione levels were measured using the commercial GSH/GSSG Quantification Kit (Dojindo) according to the manufacturer's instructions.

Techniques: Inhibition

Rg3@PACVs attenuate oxidative stress and mitochondrial dysfunction in hypoxia/reoxygenation (H/R)-injured lung epithelial cells (A) Reactive oxygen species (ROS) levels detected by flow cytometry using DCFH-DA probe. (B) Intracellular total superoxide dismutase (T-SOD) activity. (C) Malondialdehyde (MDA) content. (D) Glutathione/oxidized glutathione (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and heme oxygenase-1 (HO-1, red). Nuclei counterstained with DAPI (blue). Scale bars, 100 μm. (F) Mitochondrial membrane potential assessed by JC-1 staining. (G) Super-resolution microscopy of mitochondrial ultrastructure (red) in epithelial cells. Lower panel shows magnified views (∗ represents damaged mitochondria, Scale bars, 5 μm). (H–L) Mitochondrial respiration in BEAS-2B cells under different stimulation conditions was assessed by measuring the oxygen consumption rate (OCR) using a Seahorse XF96 analyzer (H). Key parameters including basal respiration (I), maximal respiration (J), proton leak (K), and ATP production (L) were calculated. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 4 biological replicates).

Journal: Cell Reports Medicine

Article Title: Macrophage-mimetic photothermal nanotherapeutics regulate mitochondrial homeostasis and inflammatory cascades in lung ischemia-reperfusion injury

doi: 10.1016/j.xcrm.2026.102768

Figure Lengend Snippet: Rg3@PACVs attenuate oxidative stress and mitochondrial dysfunction in hypoxia/reoxygenation (H/R)-injured lung epithelial cells (A) Reactive oxygen species (ROS) levels detected by flow cytometry using DCFH-DA probe. (B) Intracellular total superoxide dismutase (T-SOD) activity. (C) Malondialdehyde (MDA) content. (D) Glutathione/oxidized glutathione (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and heme oxygenase-1 (HO-1, red). Nuclei counterstained with DAPI (blue). Scale bars, 100 μm. (F) Mitochondrial membrane potential assessed by JC-1 staining. (G) Super-resolution microscopy of mitochondrial ultrastructure (red) in epithelial cells. Lower panel shows magnified views (∗ represents damaged mitochondria, Scale bars, 5 μm). (H–L) Mitochondrial respiration in BEAS-2B cells under different stimulation conditions was assessed by measuring the oxygen consumption rate (OCR) using a Seahorse XF96 analyzer (H). Key parameters including basal respiration (I), maximal respiration (J), proton leak (K), and ATP production (L) were calculated. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. control; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. H/R; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. H/R + Rg3@PACVs; n = 4 biological replicates).

Article Snippet: Oxidative stress markers were quantified with assay kits: MDA (G4300 and G4302, Servicebio), GSH/GSSG (G4304, Servicebio), T-SOD (G4306, Servicebio) and ATP (G4309, Servicebio).

Techniques: Flow Cytometry, Activity Assay, Immunofluorescence, Membrane, Staining, Super-Resolution Microscopy, Control

Rg3@PACVs alleviate oxidative stress impairment and mitochondrial damage in rat lung ischemia-reperfusion injury (IRI) (A–D) Assessment of oxidative stress in lung tissue. (A) Reactive oxygen species (ROS) fluorescence staining (red). Scale bars, 50 μm. (B) Total superoxide dismutase (T-SOD) activity. (C) Glutathione/oxidized glutathione (GSH/GSSG) ratio. (D) Malondialdehyde (MDA) content. (E and F) Evaluation of energy metabolism alterations. (E) Adenosine triphosphate (ATP) levels measured by biochemical assay. (F) Transmission electron microscopy (TEM) analysis of mitochondrial ultrastructural changes. Scale bars, 1 μm. (G) Immunofluorescence co-localization analysis of epithelial marker E-cadherin (red) and damage marker high-mobility group box 1 (HMGB1, green) in lung tissue. Nuclei counterstained DAPI (blue). Scale bars, 50 μm. (H and I) Preparation of single-cell suspension from lung tissue and detection of changes in the proportion of neutrophils (H) and classical monocytes (I) using flow cytometry. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. Sham; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. IRI; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. IRI+Rg3@PACVs; n = 5 biological replicates for B–E and n = 3 biological replicates for H and I).

Journal: Cell Reports Medicine

Article Title: Macrophage-mimetic photothermal nanotherapeutics regulate mitochondrial homeostasis and inflammatory cascades in lung ischemia-reperfusion injury

doi: 10.1016/j.xcrm.2026.102768

Figure Lengend Snippet: Rg3@PACVs alleviate oxidative stress impairment and mitochondrial damage in rat lung ischemia-reperfusion injury (IRI) (A–D) Assessment of oxidative stress in lung tissue. (A) Reactive oxygen species (ROS) fluorescence staining (red). Scale bars, 50 μm. (B) Total superoxide dismutase (T-SOD) activity. (C) Glutathione/oxidized glutathione (GSH/GSSG) ratio. (D) Malondialdehyde (MDA) content. (E and F) Evaluation of energy metabolism alterations. (E) Adenosine triphosphate (ATP) levels measured by biochemical assay. (F) Transmission electron microscopy (TEM) analysis of mitochondrial ultrastructural changes. Scale bars, 1 μm. (G) Immunofluorescence co-localization analysis of epithelial marker E-cadherin (red) and damage marker high-mobility group box 1 (HMGB1, green) in lung tissue. Nuclei counterstained DAPI (blue). Scale bars, 50 μm. (H and I) Preparation of single-cell suspension from lung tissue and detection of changes in the proportion of neutrophils (H) and classical monocytes (I) using flow cytometry. Data are presented as the mean ± SD and analyzed using one-way ANOVA with Tukey’s post hoc test ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. Sham; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. IRI; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. IRI+Rg3@PACVs; n = 5 biological replicates for B–E and n = 3 biological replicates for H and I).

Article Snippet: Oxidative stress markers were quantified with assay kits: MDA (G4300 and G4302, Servicebio), GSH/GSSG (G4304, Servicebio), T-SOD (G4306, Servicebio) and ATP (G4309, Servicebio).

Techniques: Fluorescence, Staining, Activity Assay, Transmission Assay, Electron Microscopy, Immunofluorescence, Marker, Single Cell, Suspension, Flow Cytometry