Journal: Scientific Reports

Article Title: AOH induces oxidative stress and DNA damage in ovarian cancer cells via modulation of GPER1 and HIF1α/PI3K/CLDNs signaling pathway

doi: 10.1038/s41598-025-26042-9

Figure Lengend Snippet: AOH induces oxidative stress in OVCAR3 cells. ( a ) Oxidative stress: The analysis of the generation of ROS with flow cytometry. ( b ) Representative results of flow cytometry. ( c ) SOD2 protein expression: Expression of SOD2 was evaluated with Western Blot, the representative results. ( d ) SOD1, SOD2, PARP1 and XRCC1 gene expression: Relative expression of SOD1, SOD2 , PARP1 and XRCC1 . ( e ) DNA damage: evaluated with flow cytometry (ATM and H2A.X activation). ( f ) Representative results of flow cytometry. ( g ) p-H2A.X, PARP1 and cleaved-PARP1 protein expression: Expression of phospho-H2A.X, PARP1 and cleaved—PARP1 evaluated with Western Blot, the representative results. ( h ) Confocal microscopy: Bar graph showing the difference in normalized mean fluorescence intensity signal. ( i ) Immunofluorescent staining of phospho-H2A.X (green) and nucleus (orange, NucSpot 555 / 570). All the results are expressed as mean ± SE. One-way ANOVA with Bonferroni correction was used for statistical analysis. P < 0.05 was considered statistically significant, * p < 0.05, ** p < 0.01, *** p < 0.001. AOH alternariol, G15 selective GPER1 receptor antagonist, G1 selective GPER1 agonist, SOD1 Superoxide dismutase 1, SOD2 Superoxide dismutase 2, PARP1 Poly [ADP-ribose] polymerase 1, XRCC1 X-ray repair cross-complementing protein 1, GAPDH anti-glyceraldehyde 3-phosphate dehydrogenase, pH2AX phospho-H2A histone family member X. Cnt control (non-treated cells).

Article Snippet: Akt (#9272), phospho-Akt (Ser473) (#4060), PARP1 (#46,011), SOD1 (#4266), SOD2 (#13,141), pH2AX (#9718), CLDN1 (#13,995), CLDN3 (#83,609), PI3K (#11,889), p44/42 (#4695), phospho-p44/42 (#4370), CLDN4 (STJ114778), GPER1 (STJ192629) (St John’s Laboratory Ltd, University Way, London) antibodies were used according to the manufacturer’s recommendations.

Techniques: Flow Cytometry, Expressing, Western Blot, Gene Expression, Activation Assay, Confocal Microscopy, Fluorescence, Staining, Control

Journal: Animal Cells and Systems

Article Title: Epinephrine as a potential driver of oral lichen planus pathogenesis

doi: 10.1080/19768354.2025.2588914

Figure Lengend Snippet: High level of epinephrine induces intracellular oxidative stress in oral keratinocytes. (A) HOK-16B cells treated with PBS or epinephrine for 24 h were subjected to CellROX staining. Nuclear DAPI signal is shown in blue (left). The relative CellROX signal intensities are shown (right). Each symbol means an individual cell. (B) Immunoblot analysis of SOD2 and SESN2 in HOK-16B cells treated with epinephrine for 24 h. β ACTIN was used as loading control. (C) Relative band intensities are shown. (D) Relative mRNA expression of TNFα, IL6, and IL8 in HOK-16B cells treated with epinephrine for 3 h. ** P < 0.01; *** P < 0.001. n.s , not significant.

Article Snippet: After blocking with 5% non-fat dried milk in Tris-buffered saline with TweenTM 20 (TBST) (10 mM Tris, pH 8.0, 150 mM NaCl and 0.5% Tween 20) for 30 min, membranes were incubated with antibodies against phospho STAT3 (1:1000, Cell signaling, Danvers, MA, USA, cat no. 9145), total STAT3 (1:1000, Cell signaling, cat no. 8768), β ACTIN (1:5000, Sigma Aldrich, cat no. A5316), phospho ERK (1:1000, Thermo Fisher Scientific, Waltham, MA, USA, cat no. MA5-15174), total ERK (1:1000, Cell signaling, cat no. 4695), phospho AKT (1:1000, Cell signaling, cat no. 4058), total AKT (1:1000, Cell signaling, cat no. 9272), BCL2 (1:1000, Bioworld Technology, China, cat no. BS1511), SOD2 (1:1000, Cusabio, Houston, TX, USA, cat no. CSB-PA022398LA01HU), and SESN2 (1:1000, Abcam, Cambridge, UK, cat no. ab178518) overnight at 4°C.

Techniques: Staining, Western Blot, Control, Expressing

Journal: Frontiers in Endocrinology

Article Title: Age- and diet-dependent progression of retinal microvascular injury in GCK-MODY under metabolic stress

doi: 10.3389/fendo.2026.1744691

Figure Lengend Snippet: HFD increases retinal oxidative stress in GCK Mut at 60 weeks. (A) Retinal reactive oxygen species (ROS) production at different time points was assessed by dihydroethidium (DHE) staining. (B) Quantification of fluorescence intensity in each experimental group. (C–E) Western blot analysis of NOX2 and SOD2 expression, with representative protein bands and expression levels at 28, 40, and 60 weeks of age. Scale bar = 50 μm. n = 6 mice per group. Data are presented as mean ± SEM. *P < 0.05, **P < 0.01, ns, not significant (GCK Mut +HFD vs. WT+HFD).

Article Snippet: After blocking, membranes were incubated with primary antibodies targeting IL-1β (A16288; ABclonal), TNF-α (A11534; ABclonal), NOX2 (BM4576; BOSTER), SOD2 (PB9442; BOSTER), Bcl-2 (BA0412; BOSTER), Bax (A00183; BOSTER), Cleaved caspase-3 (A2156; ABclonal), p-S6K (AP0564; ABclonal), S6K (A4898; ABclonal), p-S6 (AP1328; ABclonal), S6 (A11874; ABclonal), and β-actin (KM9001T; Sungene).

Techniques: Staining, Fluorescence, Western Blot, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: D4F prophylaxis enables redox and energy homeostasis while preventing inflammation during hypoxia exposure.

doi: 10.1016/j.biopha.2020.111083

Figure Lengend Snippet: Fig. 5. a. Hypoxia responsive signaling cascade associated with apolipoproteins. A STRING [88] based network with colored spheres showing query proteins; medium con fidence (0.400); 20 interactors in second shell showing the links between master regulatory protein (HIF-1a); antioxidants (SOD); inflam matory cytokines (CRP) and metabolic proteins (apolipoproteins). b. Representative immu noblots and bar-graphs depicting pixel in tensities of these immunoblots in HIF-1a and MnSOD in lungs and CRP, Apo-A1 and Apo-B in plasma. HIF-1a pixel intensities in N-2241.368 ± 162.04, ND-3985.723 ± 266.038, H-8679.09 ± 309.545, HD-9264.693 ± 394.014. MnSOD pixel intensities in N-26374.56 ± 754, ND-31417.26 ± 841, H-34801.78 ± 632, HD-39799.67 ± 432. Pixel intensities of CRP in N-990.134 ± 144.87, ND-8519.572 ± 358.33, H-17471.86 ± 511.998, HD-9972.995 ± 655.713. Apo-A1 pixel intensities in N-35004.76 ± 1058.93, ND-40377.78 ± 1264, H-29687.65 ± 1199.14, HD-30482.96 ± 983. Apo-B pixel intensities in N-22390.02 ± 244, ND-23844.02 ± 739.99, H-30615 ± 855.62, HD-24528.61 ± 730.55. All results arepresented as Mean ± SEM in AU (arbitrary units). Significance represented as * (p-value<0.05). Data from three separate bio logical replicates.Equal loading control was whole gel stained with Coomassie blue (Fig. S1; Supplementary information).

Article Snippet: The following primary antibodies were used for immunoblotting: HIF-1a (Cat. # NB100105, Novus Biologicals, USA); SOD (Cat # CSB-PA022398KA01HU; Cusabio, USA); CRP (Cat # ab5086; Abcam, USA); Apo-A1 (Cat. # sc-30089, Santa Cruz, UK) and Apo-B (Cat # CSB-PA318254; Cusabio; USA).

Techniques: Western Blot, Clinical Proteomics, Control, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential autophagy power in the spinal cord and muscle of transgenic ALS mice

doi: 10.3389/fncel.2013.00234

Figure Lengend Snippet: Biochemical properties of wtSOD1 and mutSOD1 in spinal cord and muscle of ALS animal models . Western Blot (WB) analysis and Filter Retardation Assay (FRA) on proteins extracted from whole spinal cord ( A , WB; C , FRA) or quadriceps muscles ( B , WB; D , FRA) of female and male non-transgenic (NTg) mice, of mice expressing the wild type human SOD1 transgene (wtSOD1), and of mice expressing the G93A mutant form of human SOD1 (G93A-SOD1), at 8 (corresponding to presymptomatic stage, PS, in Tg G93A-SOD1 mice) or 16 weeks (corresponding to symptomatic stage, S, in Tg G93A-SOD1 mice). GAPDH was used to normalize protein loading. hSOD1, transgenic human SOD1; mSOD1, endogenous murine SOD1.

Article Snippet: The membranes were treated with a blocking solution containing 5% non-fat dried milk powder (EuroClone) in TBS-T for 1 h and then incubated with the primary antibodies: (1) home-made rabbit polyclonal anti-HSPB8 (kindly provided by Dr Jacques Landry, Quèbec, Canada, dilution 1:1000); (2) rabbit-polyclonal anti-BAG3 antibody (Abcam, Cambridge, UK; dilution 1:1000); (3) rabbit-polyclonal anti-BAG1 (Santa Cruz Biotechnology, Dallas, Texas, USA; dilution 1:1000); (4) rabbit polyclonal anti-LC3 (Sigma; dilution 1:1000); (5) rabbit polyclonal anti-SQSTM1/p62 (Abcam; dilution 1:1000); (6) rabbit-polyclonal anti-GAPDH (Santa Cruz Biotech; dilution 1:1000); (7) rabbit-polyclonal anti-SOD1 (SOD-100; StressMarq, Victoria, BC, Canada; dilution 1:1000).

Techniques: Western Blot, Transgenic Assay, Expressing, Mutagenesis

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential autophagy power in the spinal cord and muscle of transgenic ALS mice

doi: 10.3389/fncel.2013.00234

Figure Lengend Snippet: Changes in HSPB8 levels in response to wt or mutSOD1 expression in spinal cord and muscle of ALS animal models . RT-qPCRs were performed on total RNA extracted from whole spinal cord (A) or quadriceps muscles (B) of female and male non-transgenic (NTg) mice, of mice expressing the wild type human SOD1 transgene (Tg wtSOD1), and of mice expressing the G93A mutant form of human SOD1 (Tg G93A-SOD1), at 8 (corresponding to presymptomatic stage, PS, in Tg G93A-SOD1 mice) or 16 weeks (corresponding to symptomatic stage, S, in Tg G93A-SOD1 mice). Data have been normalized to the amount of GAPDH mRNA, expressed relative to the levels determined in age-matched PS female NTg mice, taken as internal reference, and expressed as fold changes. Data are mean ± SD of four independent replicates. (A) ** p < 0.01 vs. age- and sex-matched NTg mice. (B) ** p < 0.01 vs. age- and sex-matched NTg mice and Tg wt SOD1 mice; °° p < 0.01 vs. PS (8 weeks) female Tg G93A-SOD1; # p < 0.05 vs. PS (8 weeks) male Tg G93A-SOD1.

Article Snippet: The membranes were treated with a blocking solution containing 5% non-fat dried milk powder (EuroClone) in TBS-T for 1 h and then incubated with the primary antibodies: (1) home-made rabbit polyclonal anti-HSPB8 (kindly provided by Dr Jacques Landry, Quèbec, Canada, dilution 1:1000); (2) rabbit-polyclonal anti-BAG3 antibody (Abcam, Cambridge, UK; dilution 1:1000); (3) rabbit-polyclonal anti-BAG1 (Santa Cruz Biotechnology, Dallas, Texas, USA; dilution 1:1000); (4) rabbit polyclonal anti-LC3 (Sigma; dilution 1:1000); (5) rabbit polyclonal anti-SQSTM1/p62 (Abcam; dilution 1:1000); (6) rabbit-polyclonal anti-GAPDH (Santa Cruz Biotech; dilution 1:1000); (7) rabbit-polyclonal anti-SOD1 (SOD-100; StressMarq, Victoria, BC, Canada; dilution 1:1000).

Techniques: Expressing, Transgenic Assay, Mutagenesis, Mouse Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential autophagy power in the spinal cord and muscle of transgenic ALS mice

doi: 10.3389/fncel.2013.00234

Figure Lengend Snippet: BAG3 and BAG1 expression in spinal cord and muscle of ALS animal models . RT-qPCRs were performed on total RNA extracted from whole spinal cord (A,C) or quadriceps muscles (B,D) of female and male non-transgenic (NTg) mice, of mice expressing the wild type human SOD1 transgene (Tg wtSOD1), and of mice expressing the G93A mutant form of human SOD1 (Tg G93A-SOD1), at 8 (corresponding to presymptomatic stage, PS, in Tg G93A-SOD1 mice) or 16 weeks (corresponding to symptomatic stage, S, in Tg G93A-SOD1 mice). Data have been normalized to the amount of GAPDH mRNA, expressed relative to the levels determined in age-matched PS female NTg mice, taken as internal reference, and expressed as fold changes. Data are mean ± SD of four independent replicates. (A) RT-qPCR on BAG3 mRNA expression levels in whole spinal cord. ** p < 0.01 vs. age- and sex-matched NTg and Tg wtSOD1 mice; ∧ p < 0.05 vs. age- and sex-matched NTg mice; §§ p < 0.01 vs. age- and sex-matched Tg wtSOD1 mice. (B) RT-qPCR on BAG3 mRNA expression levels in quadriceps muscles. ** p < 0.01 vs. age- and sex-matched NTg and Tg wtSOD1 mice; ° p < 0.05 vs. PS (8 weeks) female Tg G93A-SOD1; ## p < 0.01 vs. PS (8 weeks) male Tg G93A-SOD1. (C) RT-qPCR on BAG1 mRNA expression levels in whole spinal cord. (D) RT-qPCR on BAG1 mRNA expression levels in quadriceps muscles. ** p < 0.01 vs. age- and sex-matched NTg and Tg wtSOD1 mice; ° p < 0.05 vs. sex-matched PS (8 weeks) Tg G93A-SOD1. (E) BAG3:BAG1 relative ratio of mRNA expression levels in whole spinal cord. Data represent variations of the relative levels of BAG3 and BAG1 normalized over the relative BAG3 and BAG1 levels of age-matched PS (8 weeks) female NTg mice (taken as internal reference, see Materials and Methods for details). ** p < 0.01 vs. age- and sex-matched NTg and Tg wtSOD1 mice; ## p < 0.01 vs. PS (8 weeks) male Tg G93A-SOD1. (F) BAG3:BAG1 relative ratio of mRNA expression levels in quadriceps muscles. Data have been calculated as in (E) . * p < 0.05 and ** p < 0.01 vs. age- and sex-matched NTg and Tg wtSOD1 mice; ° p < 0.05 vs. PS (8 weeks) male Tg G93A-SOD1.

Article Snippet: The membranes were treated with a blocking solution containing 5% non-fat dried milk powder (EuroClone) in TBS-T for 1 h and then incubated with the primary antibodies: (1) home-made rabbit polyclonal anti-HSPB8 (kindly provided by Dr Jacques Landry, Quèbec, Canada, dilution 1:1000); (2) rabbit-polyclonal anti-BAG3 antibody (Abcam, Cambridge, UK; dilution 1:1000); (3) rabbit-polyclonal anti-BAG1 (Santa Cruz Biotechnology, Dallas, Texas, USA; dilution 1:1000); (4) rabbit polyclonal anti-LC3 (Sigma; dilution 1:1000); (5) rabbit polyclonal anti-SQSTM1/p62 (Abcam; dilution 1:1000); (6) rabbit-polyclonal anti-GAPDH (Santa Cruz Biotech; dilution 1:1000); (7) rabbit-polyclonal anti-SOD1 (SOD-100; StressMarq, Victoria, BC, Canada; dilution 1:1000).

Techniques: Expressing, Transgenic Assay, Mutagenesis, Quantitative RT-PCR, Mouse Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential autophagy power in the spinal cord and muscle of transgenic ALS mice

doi: 10.3389/fncel.2013.00234

Figure Lengend Snippet: HSPB8-BAG1-BAG3 protein levels in spinal cord and muscle of ALS animal models . Western Blot (WB) analysis on proteins extracted from whole spinal cord (A) or quadriceps muscles (B) of female and male non-transgenic (NTg) mice, of mice expressing the wild type human SOD1 transgene (Tg wtSOD1), and of mice expressing the G93A mutant form of human SOD1 (Tg G93A-SOD1), at 8 (corresponding to presymptomatic stage, PS, in Tg G93A-SOD1 mice) or 16 weeks (corresponding to symptomatic stage, S, in Tg G93A-SOD1 mice). GAPDH was used to normalize protein loading.

Article Snippet: The membranes were treated with a blocking solution containing 5% non-fat dried milk powder (EuroClone) in TBS-T for 1 h and then incubated with the primary antibodies: (1) home-made rabbit polyclonal anti-HSPB8 (kindly provided by Dr Jacques Landry, Quèbec, Canada, dilution 1:1000); (2) rabbit-polyclonal anti-BAG3 antibody (Abcam, Cambridge, UK; dilution 1:1000); (3) rabbit-polyclonal anti-BAG1 (Santa Cruz Biotechnology, Dallas, Texas, USA; dilution 1:1000); (4) rabbit polyclonal anti-LC3 (Sigma; dilution 1:1000); (5) rabbit polyclonal anti-SQSTM1/p62 (Abcam; dilution 1:1000); (6) rabbit-polyclonal anti-GAPDH (Santa Cruz Biotech; dilution 1:1000); (7) rabbit-polyclonal anti-SOD1 (SOD-100; StressMarq, Victoria, BC, Canada; dilution 1:1000).

Techniques: Western Blot, Transgenic Assay, Expressing, Mutagenesis

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential autophagy power in the spinal cord and muscle of transgenic ALS mice

doi: 10.3389/fncel.2013.00234

Figure Lengend Snippet: LC3 and p62 expression in spinal cord and muscle of ALS animal models . RT-qPCRs were performed on total RNA extracted from whole spinal cord (A,C) or quadriceps muscles (B,D) of female and male non-transgenic (NTg) mice, of mice expressing the wild type human SOD1 transgene (Tg wtSOD1), and of mice expressing the G93A mutant form of human SOD1 (Tg G93A-SOD1), at 8 (corresponding to presymptomatic stage, PS, in Tg G93A-SOD1 mice) or 16 weeks (corresponding to symptomatic stage, S, in Tg G93A-SOD1 mice). Data have been normalized to the amount of GAPDH mRNA, expressed relative to the levels determined in age-matched PS female NTg mice, taken as internal reference, and expressed as fold changes. Data are mean ± SD of four independent replicates. (A) RT-qPCR on LC3 mRNA expression levels in whole spinal cord. ** p < 0.01 vs. age- and sex-matched NTg and Tg wtSOD1 mice. (B) RT-qPCR on LC3 mRNA expression levels in quadriceps muscles. ∧ p < 0.05 vs. age- and sex-matched NTg mice; §§ p < 0.01 vs. age- and sex-matched Tg wtSOD1 mice. (C) RT-qPCR on p62 mRNA expression levels in whole spinal cord. (D) RT-qPCR on p62 mRNA expression levels in quadriceps muscles. ** p < 0.01 vs. age- and sex-matched NTg and Tg wtSOD1 mice; °° p < 0.01 vs. PS female (8 weeks) Tg G93A-SOD1.

Article Snippet: The membranes were treated with a blocking solution containing 5% non-fat dried milk powder (EuroClone) in TBS-T for 1 h and then incubated with the primary antibodies: (1) home-made rabbit polyclonal anti-HSPB8 (kindly provided by Dr Jacques Landry, Quèbec, Canada, dilution 1:1000); (2) rabbit-polyclonal anti-BAG3 antibody (Abcam, Cambridge, UK; dilution 1:1000); (3) rabbit-polyclonal anti-BAG1 (Santa Cruz Biotechnology, Dallas, Texas, USA; dilution 1:1000); (4) rabbit polyclonal anti-LC3 (Sigma; dilution 1:1000); (5) rabbit polyclonal anti-SQSTM1/p62 (Abcam; dilution 1:1000); (6) rabbit-polyclonal anti-GAPDH (Santa Cruz Biotech; dilution 1:1000); (7) rabbit-polyclonal anti-SOD1 (SOD-100; StressMarq, Victoria, BC, Canada; dilution 1:1000).

Techniques: Expressing, Transgenic Assay, Mutagenesis, Quantitative RT-PCR, Mouse Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential autophagy power in the spinal cord and muscle of transgenic ALS mice

doi: 10.3389/fncel.2013.00234

Figure Lengend Snippet: Autophagic-markers (p62 and LC-3) protein levels in spinal cord and muscle of ALS animal models . Western Blot (WB) analysis on proteins extracted from whole spinal cord (A) or quadriceps muscles (B) of female and male non-transgenic (NTg) mice, of mice expressing the wild type human SOD1 transgene (Tg wtSOD1), and of mice expressing the G93A mutant form of human SOD1 (Tg G93A-SOD1), at 8 (corresponding to presymptomatic stage, PS, in Tg G93A-SOD1 mice) or 16 weeks (corresponding to symptomatic stage, S, in Tg G93A-SOD1 mice). GAPDH was used to normalize protein loading.

Article Snippet: The membranes were treated with a blocking solution containing 5% non-fat dried milk powder (EuroClone) in TBS-T for 1 h and then incubated with the primary antibodies: (1) home-made rabbit polyclonal anti-HSPB8 (kindly provided by Dr Jacques Landry, Quèbec, Canada, dilution 1:1000); (2) rabbit-polyclonal anti-BAG3 antibody (Abcam, Cambridge, UK; dilution 1:1000); (3) rabbit-polyclonal anti-BAG1 (Santa Cruz Biotechnology, Dallas, Texas, USA; dilution 1:1000); (4) rabbit polyclonal anti-LC3 (Sigma; dilution 1:1000); (5) rabbit polyclonal anti-SQSTM1/p62 (Abcam; dilution 1:1000); (6) rabbit-polyclonal anti-GAPDH (Santa Cruz Biotech; dilution 1:1000); (7) rabbit-polyclonal anti-SOD1 (SOD-100; StressMarq, Victoria, BC, Canada; dilution 1:1000).

Techniques: Western Blot, Transgenic Assay, Expressing, Mutagenesis

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Age-related reduction in the expression of FOXO transcription factors and correlations with intervertebral disc degeneration

doi: 10.1002/jor.23583

Figure Lengend Snippet: Changes in SESN3 and SOD2 expression in mouse intervertebral discs (IVD) during aging. A) Representative images of SESN3 and SOD2 immunohistochemical staining in lumbar IVD from 6, 12, 24, and 36-month-old mice (n=5 per group). Magnification bar = 50 μm. B) Quantification of SESN3 and SOD2 immunopositive cells in the nucleus pulposus of lumbar IVD from 6, 12, 24, and 36-month-old mice (n=5 per group). All values are mean ± 95% CI.

Article Snippet: The sections were blocked with 10% goat serum for 1 hour at room temperature followed by an overnight incubation with primary antibodies against FOXO1 (1:250, Abcam, ab70382), FOXO3 (1:500, Abcam, ab70315), FOXO4 (1:500, Abcam, ab63254), SOD2 (1:500, StressMarq Biosciences, SPC-118), SESN3 (1:100, Proteintech, 24532-1-AP) at 4°C.

Techniques: Expressing, Immunohistochemical staining, Staining