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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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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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rabbit anti hiv 1 gag p24 polyclonal antibody  (Sino Biological)
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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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (GSH/GSSG) ratio. (E) Immunofluorescence detection of nuclear-factor-erythroid-2-related factor 2 (NRF-2, green) and <t>heme</t> <t>oxygenase-1</t> (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 (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: Anti-Heme Oxygenase 1 Rabbit pAb , Servicebio , Cat# GB115713.

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