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fluorescent probe sirhonox 1  (GORYO Chemical)


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    GORYO Chemical fluorescent probe sirhonox 1
    TRIM59 mitigates ferroptosis in vitro . Neuronal cells were transfected with Lv-shTRIM59 or Lv-TRIM59 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B, C) Transfection efficiency was confirmed through western blot analysis (n = 3); (D) Cell viability was determined using CCK-8 assay (n = 6); (E, F) Neuronal survival was examined by Calcein-AM/PI double staining (n = 3); (G-K) ROS, Fe 2+ , MDA, and GSH levels were measured (n = 3); (L, M) Fe 2+ accumulation was visualized <t>with</t> <t>SiRhoNox-1</t> staining (n = 3); (N) Ferroptosis-related protein expression was assessed by western blot (n = 3); (O, P) Mitochondrial ROS accumulation was measured using MitoSOX immunofluorescence and corresponding statistical analysis (n = 3); (Q, R) Mitochondrial membrane potential was assessed via JC-1 staining, with quantification of JC-1 aggregates and monomers (n = 3); (S) Representative TEM images of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Fluorescent Probe Sirhonox 1, supplied by GORYO Chemical, used in various techniques. Bioz Stars score: 95/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/fluorescent+probe+sirhonox+1/FerroFarRed/pmc13068617-88-12-16
    Average 95 stars, based on 93 article reviews
    fluorescent probe sirhonox 1 - by Bioz Stars, 2026-09
    95/100 stars

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    1) Product Images from "TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2"

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2

    Journal: Journal of Orthopaedic Translation

    doi: 10.1016/j.jot.2026.101070

    TRIM59 mitigates ferroptosis in vitro . Neuronal cells were transfected with Lv-shTRIM59 or Lv-TRIM59 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B, C) Transfection efficiency was confirmed through western blot analysis (n = 3); (D) Cell viability was determined using CCK-8 assay (n = 6); (E, F) Neuronal survival was examined by Calcein-AM/PI double staining (n = 3); (G-K) ROS, Fe 2+ , MDA, and GSH levels were measured (n = 3); (L, M) Fe 2+ accumulation was visualized with SiRhoNox-1 staining (n = 3); (N) Ferroptosis-related protein expression was assessed by western blot (n = 3); (O, P) Mitochondrial ROS accumulation was measured using MitoSOX immunofluorescence and corresponding statistical analysis (n = 3); (Q, R) Mitochondrial membrane potential was assessed via JC-1 staining, with quantification of JC-1 aggregates and monomers (n = 3); (S) Representative TEM images of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: TRIM59 mitigates ferroptosis in vitro . Neuronal cells were transfected with Lv-shTRIM59 or Lv-TRIM59 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B, C) Transfection efficiency was confirmed through western blot analysis (n = 3); (D) Cell viability was determined using CCK-8 assay (n = 6); (E, F) Neuronal survival was examined by Calcein-AM/PI double staining (n = 3); (G-K) ROS, Fe 2+ , MDA, and GSH levels were measured (n = 3); (L, M) Fe 2+ accumulation was visualized with SiRhoNox-1 staining (n = 3); (N) Ferroptosis-related protein expression was assessed by western blot (n = 3); (O, P) Mitochondrial ROS accumulation was measured using MitoSOX immunofluorescence and corresponding statistical analysis (n = 3); (Q, R) Mitochondrial membrane potential was assessed via JC-1 staining, with quantification of JC-1 aggregates and monomers (n = 3); (S) Representative TEM images of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: In Vitro, Transfection, Western Blot, CCK-8 Assay, Double Staining, Staining, Expressing, Immunofluorescence, Membrane

    Lv-shANXA2 mitigates the ferroptosis induced by TRIM59 knockdown. Neuronal cells were transfected with either Lv-shANXA2 or Lv-ANXA2 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B) Cell viability assessment via CCK-8 assay (n = 6); (C, D) Neuronal survival evaluation through Calcein-AM/PI staining (n = 3); (E, F) Intracellular ROS quantification by flow cytometry with statistical analysis (n = 3); (G-I) Measurement of Fe 2+ , MDA, and GSH levels in neuronal cells (n = 3); (J, K) Fe 2+ accumulation detection using SiRhoNox-1 staining (n = 3); (L) Western blot examination of ferroptosis-associated protein expression in neuronal cells (n = 3); (M, N) Mitochondrial ROS detection and quantification via MitoSOX immunofluorescence in neuronal cells (n = 3); (O, P) Mitochondrial membrane potential assessment through JC-1 staining, including JC-1 aggregate-to-monomer ratio quantification (n = 3); (Q) Transmission electron microscopy visualization of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: Lv-shANXA2 mitigates the ferroptosis induced by TRIM59 knockdown. Neuronal cells were transfected with either Lv-shANXA2 or Lv-ANXA2 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B) Cell viability assessment via CCK-8 assay (n = 6); (C, D) Neuronal survival evaluation through Calcein-AM/PI staining (n = 3); (E, F) Intracellular ROS quantification by flow cytometry with statistical analysis (n = 3); (G-I) Measurement of Fe 2+ , MDA, and GSH levels in neuronal cells (n = 3); (J, K) Fe 2+ accumulation detection using SiRhoNox-1 staining (n = 3); (L) Western blot examination of ferroptosis-associated protein expression in neuronal cells (n = 3); (M, N) Mitochondrial ROS detection and quantification via MitoSOX immunofluorescence in neuronal cells (n = 3); (O, P) Mitochondrial membrane potential assessment through JC-1 staining, including JC-1 aggregate-to-monomer ratio quantification (n = 3); (Q) Transmission electron microscopy visualization of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Knockdown, Transfection, CCK-8 Assay, Staining, Flow Cytometry, Western Blot, Expressing, Immunofluorescence, Membrane, Transmission Assay, Electron Microscopy

    Related Articles

    In Vitro:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Transfection:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Western Blot:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    CCK-8 Assay:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Double Staining:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Staining:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Expressing:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Immunofluorescence:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Membrane:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Knockdown:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Flow Cytometry:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Transmission Assay:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.

    Electron Microscopy:

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2
    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ]. .. C

    Article Title: Athlete-derived extracellular vesicles protect against spinal cord injury via inhibition of neuronal ferroptosis
    Article Snippet: Labile iron (Fe 2+ ) was quantified using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical, Japan) following established protocols ( ).. Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.Cells were cultured on confocal dishes and incubated with 5 μM SiRhoNox-1 for 1 hour at 37°C.



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    fluorescent probe sirhonox 1  (GORYO Chemical)
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    GORYO Chemical fluorescent probe sirhonox 1
    TRIM59 mitigates ferroptosis in vitro . Neuronal cells were transfected with Lv-shTRIM59 or Lv-TRIM59 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B, C) Transfection efficiency was confirmed through western blot analysis (n = 3); (D) Cell viability was determined using CCK-8 assay (n = 6); (E, F) Neuronal survival was examined by Calcein-AM/PI double staining (n = 3); (G-K) ROS, Fe 2+ , MDA, and GSH levels were measured (n = 3); (L, M) Fe 2+ accumulation was visualized <t>with</t> <t>SiRhoNox-1</t> staining (n = 3); (N) Ferroptosis-related protein expression was assessed by western blot (n = 3); (O, P) Mitochondrial ROS accumulation was measured using MitoSOX immunofluorescence and corresponding statistical analysis (n = 3); (Q, R) Mitochondrial membrane potential was assessed via JC-1 staining, with quantification of JC-1 aggregates and monomers (n = 3); (S) Representative TEM images of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
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    TRIM59 mitigates ferroptosis in vitro . Neuronal cells were transfected with Lv-shTRIM59 or Lv-TRIM59 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B, C) Transfection efficiency was confirmed through western blot analysis (n = 3); (D) Cell viability was determined using CCK-8 assay (n = 6); (E, F) Neuronal survival was examined by Calcein-AM/PI double staining (n = 3); (G-K) ROS, Fe 2+ , MDA, and GSH levels were measured (n = 3); (L, M) Fe 2+ accumulation was visualized <t>with</t> <t>SiRhoNox-1</t> staining (n = 3); (N) Ferroptosis-related protein expression was assessed by western blot (n = 3); (O, P) Mitochondrial ROS accumulation was measured using MitoSOX immunofluorescence and corresponding statistical analysis (n = 3); (Q, R) Mitochondrial membrane potential was assessed via JC-1 staining, with quantification of JC-1 aggregates and monomers (n = 3); (S) Representative TEM images of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Ferrofarred Known Sirhonox 1 Fluorescent Probe, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TRIM59 mitigates ferroptosis in vitro . Neuronal cells were transfected with Lv-shTRIM59 or Lv-TRIM59 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B, C) Transfection efficiency was confirmed through western blot analysis (n = 3); (D) Cell viability was determined using CCK-8 assay (n = 6); (E, F) Neuronal survival was examined by Calcein-AM/PI double staining (n = 3); (G-K) ROS, Fe 2+ , MDA, and GSH levels were measured (n = 3); (L, M) Fe 2+ accumulation was visualized with SiRhoNox-1 staining (n = 3); (N) Ferroptosis-related protein expression was assessed by western blot (n = 3); (O, P) Mitochondrial ROS accumulation was measured using MitoSOX immunofluorescence and corresponding statistical analysis (n = 3); (Q, R) Mitochondrial membrane potential was assessed via JC-1 staining, with quantification of JC-1 aggregates and monomers (n = 3); (S) Representative TEM images of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: Journal of Orthopaedic Translation

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2

    doi: 10.1016/j.jot.2026.101070

    Figure Lengend Snippet: TRIM59 mitigates ferroptosis in vitro . Neuronal cells were transfected with Lv-shTRIM59 or Lv-TRIM59 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B, C) Transfection efficiency was confirmed through western blot analysis (n = 3); (D) Cell viability was determined using CCK-8 assay (n = 6); (E, F) Neuronal survival was examined by Calcein-AM/PI double staining (n = 3); (G-K) ROS, Fe 2+ , MDA, and GSH levels were measured (n = 3); (L, M) Fe 2+ accumulation was visualized with SiRhoNox-1 staining (n = 3); (N) Ferroptosis-related protein expression was assessed by western blot (n = 3); (O, P) Mitochondrial ROS accumulation was measured using MitoSOX immunofluorescence and corresponding statistical analysis (n = 3); (Q, R) Mitochondrial membrane potential was assessed via JC-1 staining, with quantification of JC-1 aggregates and monomers (n = 3); (S) Representative TEM images of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ].

    Techniques: In Vitro, Transfection, Western Blot, CCK-8 Assay, Double Staining, Staining, Expressing, Immunofluorescence, Membrane

    Lv-shANXA2 mitigates the ferroptosis induced by TRIM59 knockdown. Neuronal cells were transfected with either Lv-shANXA2 or Lv-ANXA2 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B) Cell viability assessment via CCK-8 assay (n = 6); (C, D) Neuronal survival evaluation through Calcein-AM/PI staining (n = 3); (E, F) Intracellular ROS quantification by flow cytometry with statistical analysis (n = 3); (G-I) Measurement of Fe 2+ , MDA, and GSH levels in neuronal cells (n = 3); (J, K) Fe 2+ accumulation detection using SiRhoNox-1 staining (n = 3); (L) Western blot examination of ferroptosis-associated protein expression in neuronal cells (n = 3); (M, N) Mitochondrial ROS detection and quantification via MitoSOX immunofluorescence in neuronal cells (n = 3); (O, P) Mitochondrial membrane potential assessment through JC-1 staining, including JC-1 aggregate-to-monomer ratio quantification (n = 3); (Q) Transmission electron microscopy visualization of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: Journal of Orthopaedic Translation

    Article Title: TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2

    doi: 10.1016/j.jot.2026.101070

    Figure Lengend Snippet: Lv-shANXA2 mitigates the ferroptosis induced by TRIM59 knockdown. Neuronal cells were transfected with either Lv-shANXA2 or Lv-ANXA2 and treated with Erastin (5 μM) for 6 h. (A) Experimental group specifications; (B) Cell viability assessment via CCK-8 assay (n = 6); (C, D) Neuronal survival evaluation through Calcein-AM/PI staining (n = 3); (E, F) Intracellular ROS quantification by flow cytometry with statistical analysis (n = 3); (G-I) Measurement of Fe 2+ , MDA, and GSH levels in neuronal cells (n = 3); (J, K) Fe 2+ accumulation detection using SiRhoNox-1 staining (n = 3); (L) Western blot examination of ferroptosis-associated protein expression in neuronal cells (n = 3); (M, N) Mitochondrial ROS detection and quantification via MitoSOX immunofluorescence in neuronal cells (n = 3); (O, P) Mitochondrial membrane potential assessment through JC-1 staining, including JC-1 aggregate-to-monomer ratio quantification (n = 3); (Q) Transmission electron microscopy visualization of mitochondrial ultrastructure in neuronal cells. ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Intracellular labile iron pool (Fe 2+ ) levels were detected using the fluorescent probe SiRhoNox-1 (FerroFarRed, Goryo Chemical Japan), as previously described [ ].

    Techniques: Knockdown, Transfection, CCK-8 Assay, Staining, Flow Cytometry, Western Blot, Expressing, Immunofluorescence, Membrane, Transmission Assay, Electron Microscopy