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Effects of SESN2 on neuronal survival and oxidative stress markers in an in vitro SAH model. (A) Immunofluorescence images showing ROS production detected <t>by</t> <t>DCFH‐DA</t> staining (green) with DAPI nuclear staining (blue) and merged views across experimental groups. Scale bars = 20 μm. (B) Corresponding quantification of ROS levels in each experimental group, n = 3 per group. (C) Cell viability of HT22 cells assessed by CCK‐8 assay, n = 6 per group. (D, E) Quantitative analyses of SOD activity and MDA content, n = 6 per group. (F) Western blot analysis and quantitative analysis of SESN2 (G), GPX4 (H), SLC7A11 (I) and Nrf2 (K), n = 6 per group. (J, L) Western blot analysis and quantification of nuclear Nrf2, n = 6 per group. Data indicated as mean ± SD. ## p < 0.01, ### p < 0.001 vs. Sham group, ** p < 0.01 vs. Hemin group.
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Indocyanine green-loaded M2 macrophage-derived exosomes (ICG@M2-exos) plus near-infrared (NIR) irradiation exhibits a time-dependent dual role in reactive oxygen species <t>(ROS)</t> regulation. (A) A dose- and time-dependent increase in <t>intracellular</t> ROS levels was detected upon H 2 O 2 treatment (0.1 to 1.0 mM, 1 to 6 h). The 0.5 mM, 4-h condition (indicated) was chosen for subsequent studies. (B) Corresponding cell viability assessment for the selected condition, verifying a suitable window for intervention studies. (C) Fluorescence images showing intracellular ROS levels (green) under different conditions. (D) Quantitative fluorescence intensity analysis. Values are means ± SD ( n = 3). *** P < 0.001. a.u., arbitrary units.
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Indocyanine green-loaded M2 macrophage-derived exosomes (ICG@M2-exos) plus near-infrared (NIR) irradiation exhibits a time-dependent dual role in reactive oxygen species <t>(ROS)</t> regulation. (A) A dose- and time-dependent increase in <t>intracellular</t> ROS levels was detected upon H 2 O 2 treatment (0.1 to 1.0 mM, 1 to 6 h). The 0.5 mM, 4-h condition (indicated) was chosen for subsequent studies. (B) Corresponding cell viability assessment for the selected condition, verifying a suitable window for intervention studies. (C) Fluorescence images showing intracellular ROS levels (green) under different conditions. (D) Quantitative fluorescence intensity analysis. Values are means ± SD ( n = 3). *** P < 0.001. a.u., arbitrary units.
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Effects of SESN2 on neuronal survival and oxidative stress markers in an in vitro SAH model. (A) Immunofluorescence images showing ROS production detected by DCFH‐DA staining (green) with DAPI nuclear staining (blue) and merged views across experimental groups. Scale bars = 20 μm. (B) Corresponding quantification of ROS levels in each experimental group, n = 3 per group. (C) Cell viability of HT22 cells assessed by CCK‐8 assay, n = 6 per group. (D, E) Quantitative analyses of SOD activity and MDA content, n = 6 per group. (F) Western blot analysis and quantitative analysis of SESN2 (G), GPX4 (H), SLC7A11 (I) and Nrf2 (K), n = 6 per group. (J, L) Western blot analysis and quantification of nuclear Nrf2, n = 6 per group. Data indicated as mean ± SD. ## p < 0.01, ### p < 0.001 vs. Sham group, ** p < 0.01 vs. Hemin group.

Journal: CNS Neuroscience & Therapeutics

Article Title: Sestrin2 Mitigates Neuronal Ferroptosis Following Subarachnoid Hemorrhage via Orchestration of the AMPK / PGC1α /Nrf2 Signaling Axis

doi: 10.1002/cns.70908

Figure Lengend Snippet: Effects of SESN2 on neuronal survival and oxidative stress markers in an in vitro SAH model. (A) Immunofluorescence images showing ROS production detected by DCFH‐DA staining (green) with DAPI nuclear staining (blue) and merged views across experimental groups. Scale bars = 20 μm. (B) Corresponding quantification of ROS levels in each experimental group, n = 3 per group. (C) Cell viability of HT22 cells assessed by CCK‐8 assay, n = 6 per group. (D, E) Quantitative analyses of SOD activity and MDA content, n = 6 per group. (F) Western blot analysis and quantitative analysis of SESN2 (G), GPX4 (H), SLC7A11 (I) and Nrf2 (K), n = 6 per group. (J, L) Western blot analysis and quantification of nuclear Nrf2, n = 6 per group. Data indicated as mean ± SD. ## p < 0.01, ### p < 0.001 vs. Sham group, ** p < 0.01 vs. Hemin group.

Article Snippet: For intracellular ROS detection, HT22 cells were incubated with DCFH‐DA (10 μM, MCE) at 37°C for 30 min. Fluorescent signals were detected using excitation at 488 nm and emission was measured at 525 nm wavelength with data presented as relative fluorescence units per mg protein, normalized to sham controls.

Techniques: In Vitro, Immunofluorescence, Staining, CCK-8 Assay, Activity Assay, Western Blot

Indocyanine green-loaded M2 macrophage-derived exosomes (ICG@M2-exos) plus near-infrared (NIR) irradiation exhibits a time-dependent dual role in reactive oxygen species (ROS) regulation. (A) A dose- and time-dependent increase in intracellular ROS levels was detected upon H 2 O 2 treatment (0.1 to 1.0 mM, 1 to 6 h). The 0.5 mM, 4-h condition (indicated) was chosen for subsequent studies. (B) Corresponding cell viability assessment for the selected condition, verifying a suitable window for intervention studies. (C) Fluorescence images showing intracellular ROS levels (green) under different conditions. (D) Quantitative fluorescence intensity analysis. Values are means ± SD ( n = 3). *** P < 0.001. a.u., arbitrary units.

Journal: Research

Article Title: NIR-Activated ICG-Loaded M2 Macrophage Exosomes Ameliorate Periodontitis via Targeting Infection Inflammation and Oxidative Stress

doi: 10.34133/research.1207

Figure Lengend Snippet: Indocyanine green-loaded M2 macrophage-derived exosomes (ICG@M2-exos) plus near-infrared (NIR) irradiation exhibits a time-dependent dual role in reactive oxygen species (ROS) regulation. (A) A dose- and time-dependent increase in intracellular ROS levels was detected upon H 2 O 2 treatment (0.1 to 1.0 mM, 1 to 6 h). The 0.5 mM, 4-h condition (indicated) was chosen for subsequent studies. (B) Corresponding cell viability assessment for the selected condition, verifying a suitable window for intervention studies. (C) Fluorescence images showing intracellular ROS levels (green) under different conditions. (D) Quantitative fluorescence intensity analysis. Values are means ± SD ( n = 3). *** P < 0.001. a.u., arbitrary units.

Article Snippet: Intracellular ROS detection: Intracellular ROS levels were quantified using DCFH-DA (HY-D0940, MCE, USA).

Techniques: Derivative Assay, Irradiation, Fluorescence