Journal: BMC Neuroscience

Article Title: Sex-specific cypermethrin-induced hippocampal neurotoxicity is associated with alterations in signaling molecules for antioxidant defense, membrane integrity, apoptosis, and GABAergic integrity

doi: 10.1186/s12868-025-00988-y

Figure Lengend Snippet: GnRH ( A ), Na+/K + ATPase ( B ), PGE-2 ( C ), and COX-2 concentrations in the brains of Male and Female Wistar rats exposed to corn oil (control), low dose (6.25 mg/kg), and high dose (12.5 mg/kg) of cypermethrin, respectively. Asterisk (*) indicates significant ( P < 0.05) weight loss. Bars are mean ± SD

Article Snippet: The following kits were used: GnRH (Sandwich ELISA, Kit #MBS269200, MyBioSource), Na+/K+-ATPase (Colorimetric, Kit #E-BC-K022-M, Elabscience), PGE2 (EIA Kit #ADI-900-001, Enzo Life Sciences), COX-2 (ELISA, Kit #MBS7254655, MyBioSource).

Techniques: Control

Journal: Frontiers in Immunology

Article Title: Synergistic effects of platelet-rich fibrin and CTLA4Ig gene-transfected porcine skin on accelerating wound healing in a rat model of deep second-degree burns: a mechanistic study

doi: 10.3389/fimmu.2025.1756818

Figure Lengend Snippet: Temporal expression profiles of antioxidant enzymes CAT and SOD1. Immunofluorescence staining of (A) Catalase (CAT, red) and (B) Superoxide Dismutase 1 (SOD1, red) in wound tissues from the Vaseline group, PRF group, Pigskin group, and PRF+pig skin group at days 4, 7, 14, and 21 post-treatment. Cell nuclei are counterstained with DAPI (blue). The PRF+pig skin group shows the most pronounced and sustained enhancement in the expression of both antioxidant enzymes, particularly during the proliferative and remodeling phases (D7–D21), indicating a reinforced antioxidant defense system. Scale bar = 100 μm.

Article Snippet: After deparaffinization and antigen retrieval (similar to IHC), sections were permeabilized with 0.1% Triton X-100, blocked with 5% BSA, and incubated overnight at 4 °C with primary antibodies against CAT (1:200, Proteintech) and SOD1 (1:200, Proteintech).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Animal Models and Experimental Medicine

Article Title: Adropin modulates pancreatic cell proliferation and glutathione levels in an animal model of type 1 diabetes mellitus

doi: 10.1002/ame2.70092

Figure Lengend Snippet: Impact of adropin on superoxide dismutase expression in pancreatic β‐cells of normoglycemic and diabetic rats. (A) Immunofluorescence labeling with anti‐superoxide dismutase antibody and anti‐insulin antibodies showed expression of superoxide dismutase in pancreatic β‐cells. (B) Quantification of the histological analysis showed a significant (*** p < 0.001) decrease in superoxide dismutase distribution in pancreatic endocrine cells and a significant (**** p < 0.0001) decrease in its localization in β‐cells of rats with diabetes when compared to the normal group. n = 6. Scale bar: 50 μm. Data analysis was done using the ANOVA test.

Article Snippet: Superoxide dismutase (rabbit) (1:500) , Rockland Immunochemicals, USA.

Techniques: Expressing, Immunofluorescence, Labeling

Journal: Animal Models and Experimental Medicine

Article Title: Adropin modulates pancreatic cell proliferation and glutathione levels in an animal model of type 1 diabetes mellitus

doi: 10.1002/ame2.70092

Figure Lengend Snippet: Effect of adropin on superoxide dismutase expression in pancreatic α‐cells of normal and diabetic rats. (A) Immunofluorescence staining using anti‐superoxide dismutase and anti‐glucagon antibodies showed expression of superoxide dismutase in pancreatic α‐cells. (B) Quantification of the histological analysis showed a significant (** p < 0.01) decrease in superoxide dismutase distribution in the pancreatic endocrine cells of diabetic rats compared to the normal group. α‐Cell expression of superoxide dismutase did not change with adropin treatment among all groups. n = 6. Scale bar: 50 μm. Data analysis was done using the ANOVA test.

Article Snippet: Superoxide dismutase (rabbit) (1:500) , Rockland Immunochemicals, USA.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Animal Models and Experimental Medicine

Article Title: Adropin modulates pancreatic cell proliferation and glutathione levels in an animal model of type 1 diabetes mellitus

doi: 10.1002/ame2.70092

Figure Lengend Snippet: Effect of adropin on catalase, superoxide dismutase and total glutathione activities in the serum samples of healthy and diabetic rats. (A) Catalase activity was significantly decreased in diabetic rats compared to normal controls. Adropin slightly increased catalase in DMT group. (B) Superoxide dismutase was slightly increased in DMT compared to the diabetic treated with adropin. (C) Total glutathione was significantly raised with adropin treatment in DMT compared to DMUT. n = 4–6. Data analysis was done using the ANOVA test. * p < 0.05.

Article Snippet: Superoxide dismutase (rabbit) (1:500) , Rockland Immunochemicals, USA.

Techniques: Activity Assay

Journal: Diabetes

Article Title: Maternal Exercise-Induced SOD3 Reverses the Deleterious Effects of Maternal High-Fat Diet on Offspring Metabolism Through Stabilization of H3K4me3 and Protection Against WDR82 Carbonylation.

doi: 10.2337/db21-0706

Figure Lengend Snippet: Figure 4—Beneficial effects of maternal exercise on glucose metabolism and WDR82 carbonylation in offspring of HFD-fed dams were blocked by placenta-specific Sod3 knockout. A and B: Glucose tolerance measured at 24 weeks in Sod3f/f or Sod3/ offspring of dams that were sedentary or trained and fed chow or the HFD. Glucose area under the curve (AUC) of male (A) and female (B) offspring is shown. GTT, glucose tolerance test. Data are means ± SEM (n = 5–7/group). **P < 0.01 vs. Chow-Sod3f/f-Sed; §P < 0.01 effect of genotype; ¶P < 0.01 effect of diet. Glucose production in hepatocytes of 16-week-old male (C) and female (D) Sod3f/f or Sod3/ offspring of dams that were sedentary (Sed) or trained and fed the HFD. Data are means ± SEM (n = 3). **P < 0.01 vs. Sod3f/f-HFD-Sedentary, §P < 0.01

Article Snippet: Human SOD3 (hSOD3) cDNA was obtained from Origene (RC204156) and was cloned into p3xFLAG-CMV8 vector (E9908, Sigma-Aldrich).

Techniques: Knock-Out

Journal: Diabetes

Article Title: Maternal Exercise-Induced SOD3 Reverses the Deleterious Effects of Maternal High-Fat Diet on Offspring Metabolism Through Stabilization of H3K4me3 and Protection Against WDR82 Carbonylation.

doi: 10.2337/db21-0706

Figure Lengend Snippet: Figure 5—Effects of SOD3 on offspring glucose metabolism are distinct from NAC. A and D: Developmental system used to treat off- spring livers with recombinant SOD3 or NAC exo utero. Offspring livers were collected at 4 weeks (A) or at E13.5 (D). Glucose production in primary hepatocytes of 4-week-old male (B) and female (C) offspring of HFD-fed, saline-, SOD3-, or diethyldithiocarbamate (DETCA)- treated dams (n = 3). **P < 0.01 vs. pCPT-saline. Effects of SOD3 or NAC treatment in utero on ROS levels (E), carbonylated protein con- tent (F), WDR82 carbonylation levels (G), mRNA expression of glucose metabolism genes (H), AMPKa phosphorylation (pAMPKa) levels (I), and mRNA expression of Tet and Idh (J) in livers of E13.5 offspring of HFD-fed dams (n = 3). IP, immunoprecipitation. All data are reported as means ± SEM. **P < 0.01 vs. pCPT-saline; ***P < 0.01 vs. pCPT-saline; ****P < 0.01 vs. pCPT-saline. Statistical signifi- cance was determined by one- or two-way ANOVA, with Tukey and Bonferroni post hoc analysis.

Article Snippet: Human SOD3 (hSOD3) cDNA was obtained from Origene (RC204156) and was cloned into p3xFLAG-CMV8 vector (E9908, Sigma-Aldrich).

Techniques: Recombinant, Saline, In Utero, Expressing, Phospho-proteomics, Immunoprecipitation

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: A SOD1; B Nb1; C Nb2; D Nb3; E SOD1-Nb1 complex; F SOD1-Nb2 complex; G SOD1-Nb3 complex. The horizontal axis represents mass-to-charge ratio (m/z), and the vertical axis represents intensity, indicating the specific m/z values at peak intensities.

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques:

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: A The structure of the SOD1-Nb1 complex (PDB ID: 8K33). B The structure of the SOD1-Nb2 complex (PDB ID: 8K3A). C The structure of the SOD1-Nb3 complex (PDB ID: 8K3L). D Structural superimposition of the three nanobodies, with SOD1, Nb1, Nb2, and Nb3 shown in cyan, gray-white, yellow, and purple, respectively (Only depicted Nb1, Nb2, and Nb3 bound to one SOD1 monomer). E – G Surface electrostatic potentials of the SOD1-Nb1, SOD1-Nb2, and SOD1-Nb3 binary complexes (positive in blue, negative in red, and neutral in white). SOD1 is shown in cyan, with Cu and Zn atoms depicted as blue and orange spheres. Nb1, Nb2, and Nb3 are shown in gray-white, yellow, and purple, respectively. CDRs 1-3 of the nanobodies are highlighted in blue, green, and red, respectively.

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques:

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: SOD1 is shown in cyan, Nb1, Nb2, and Nb3 shown in gray-white, yellow, and purple, respectively with CDRs 1-3 of Nb1/Nb2/Nb3 shown in blue, green, and red, respectively. Residues involved in the interactions are depicted as sticks, with yellow dotted lines indicating hydrogen bonds and salt bridges. A Overall view and detailed views of the binding interface between SOD1 and complementarity determining regions (CDRs) of Nb1. B Overall view and detailed views of the binding interface between SOD1 and CDRs of Nb2. C Overall view and detailed views of the binding interface between SOD1 and CDRs of Nb3.

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques: Binding Assay

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: A The overall structure of SOD1, Nb1 and Nb2 complex (PDB ID: 8YAF). B The surface view of the SOD1, Nb1 and Nb2 complex. C The overall structure of SOD1, Nb1, Nb2 and Nb3 complex (PDB ID: 8YAT). D The surface view of SOD1, Nb1, Nb2 and Nb3 complex. E The structure comparison of SOD1, Nb1 and Nb2 complex with SOD1 fibril core. F The surface view of the comparison between SOD1, Nb1 and Nb2 complex with SOD1 fibril core. G The structure comparison of SOD1, Nb1, Nb2 and Nb3 complex with SOD1 fibril core. H The surface view of the comparison between SOD1, Nb1, Nb2 and Nb3 complex with SOD1 fibril core.

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques: Comparison

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: A , B Comparison of SOD1 and SOD1-nanobody binary complexes. C , D Detection of epitope binding with two nanobodies. E , F Detection of epitope binding with three nanobodies. The left panels display the full chromatogram, while the right panels provide an enlarged view of the elution peaks. SOD1, SOD1-Nb1, SOD1-Nb2, and SOD1-Nb3 are represented in black, red, blue, and green, respectively. SOD1-Nb1-Nb2, SOD1-Nb1-Nb3, SOD1-Nb2-Nb3, and SOD1-Nb1-Nb2-Nb3 are shown in purple, orange, light blue, and brown, respectively.

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques: Comparison, Binding Assay

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: A – D Binding of Nb1 to SOD1, SOD1-Nb2, SOD1-Nb3, and SOD1-Nb2-Nb3; E – H Binding of Nb2 to SOD1, SOD1-Nb1, SOD1-Nb3, and SOD1-Nb1-Nb3; I – L Binding of Nb3 to SOD1, SOD1-Nb1, SOD1-Nb2, and SOD1-Nb1-Nb2. Panels show ITC profiles with binding isotherms and corresponding thermodynamic parameters.

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques: Binding Assay

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: A , B Particle size distributions of SOD1 and its nanobody complexes by dynamic light scattering. A Size distributions for SOD1 (black), SOD1-Nb1 (red), SOD1-Nb2 (blue), and SOD1-Nb3 (green); B Size distributions for SOD1 (black), SOD1-Nb1-Nb2 (red), SOD1-Nb1-Nb3 (blue), SOD1-Nb2-Nb3 (green), and SOD1-Nb1-Nb2-Nb3 (purple). C Measurement of SOD1 enzyme activity using the Kit-WST. The activities of SOD1, SOD1-Nb1, SOD1-Nb2, and SOD1-Nb3 are shown in blue, purple, pink, and green, respectively. Error bars represent the standard error from three independent experiments (n = 3). D – G Hydrogen bond distances between Asp124 and metal-binding histidines (His46/His71) in different SOD1 conformational states. D WT SOD1, E Nb1 binding SOD1, F Nb2 binding SOD1, G Nb3 binding SOD1. H Thioflavin T (ThT) fluorescence intensities representing the filament formation of SOD1 and nanobodies binding SOD1. Error bars represent the standard error from three independent experiments (n = 3).

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques: Activity Assay, Binding Assay, Fluorescence

Journal: Communications Biology

Article Title: Structural mechanisms and insights on multiple nanobodies binding diverse SOD1 epitopes

doi: 10.1038/s42003-025-09293-0

Figure Lengend Snippet: A , B Interaction of Nb1-Nb2 tandem nanobodies with different linker lengths to SOD1. C Interaction of Nb2-Nb3 tandem nanobodies with SOD1. D Interaction of Nb1-Nb2-Nb3 tandem nanobodies with SOD1.

Article Snippet: SOD1 was labeled with Cyanine5-NHS ester iodide (MCE) at a 1:2 molar ratio at 4 °C overnight, and excess red fluorescent dye was removed using Zeba Spin Desalting columns (ThermoFisher).

Techniques:

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

Article Title: Nanoluciferase signal brightness using furimazine substrates opens bioluminescence resonance energy transfer to widefield microscopy.

doi: 10.1002/cyto.a.22870

Figure Lengend Snippet: Figure 1. Nanoluc-based bioluminescence and BRET imaging. A: HEK-293 cells show robust bioluminescence signal in the presence of Nluc-SOD1 and furimazine. In contrast, in the absence of Nluc or furimazine, no localized bioluminescence signal was detectable. Renilla luciferase tagged SOD1 protein (Rluc-SOD1) in the presence of coelenterazine showed no detectable bioluminescence signal as well. A plot of the luminescence intensity along the length of each image was measured for all three conditions. The signal-to-background ratio (S/B) of the two peaks (arrows) corresponding to the bioluminescent cells was calculated as described in the study methods. B: Scheme of the BRET positive control construct YFP-Nluc and the three-step BRET mechanism: 1) addition of the substrate furimazine producing blue light, 2) BRET of Nluc to YFP, and 3) Yellow emission light. The figure on the right shows the bioluminescence spectra properties of the fusion proteins Nluc-SOD1 (blue) and YFP-Nluc (yellow). The blue box corresponds to the band pass donor filter or Nluc channel (BP 410– 480 nm), and the yellow box corresponds to the region of the band acceptor filter or BRET channel (BP 500–550 nm). C: Bioluminescence imaging of Nluc-SOD1 and YFP-Nluc measured on acquisition bioluminescent channels (BP 410–480 nm) and (BP 500–550 nm). D: Com- parison of the net BRET for the BRET negative control and positive control constructs Nluc-SOD1 and YFP-Nluc demonstrating the robust- ness of the BRET imaging method. *** P values <0.0001. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: SOD1, Nluc and Rluc, were amplified from the SOD1 cDNA plasmid (Origene, Abingdon, UK, catalog no. SC111022), the pNL1.1 and pRL-null plasmid (Promega, Madison, WI), respectively.

Techniques: Imaging, Luciferase, Positive Control, Construct, Negative Control

Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology

Article Title: Nanoluciferase signal brightness using furimazine substrates opens bioluminescence resonance energy transfer to widefield microscopy.

doi: 10.1002/cyto.a.22870

Figure Lengend Snippet: Figure 2. SOD1 protein–protein inter-BRET quantification. A: Shown is a schematic of SOD1 dimerization. B: Scheme of the Nluc-SOD1/ SOD1-YFP and Nluc-SOD1/YFP-SOD1 constructions. C: Bioluminescence images acquired from HEK-293 cells expressing Nluc-SOD1, Nluc-SOD1/SOD1-YFP, and Nluc-SOD1/YFP-SOD1 using the Nluc channel (BP 410–480 nm) and the YFP channel (BP 500–550 nm). We created an overlay image with the following color code: Green for the Nluc channel and Red for the YFP channel. D: We calculated the net BRET for Nluc-SOD1, Nluc-SOD1/SOD1-YFP and Nluc-SOD1/YFP-SOD1 for representative cells. [Color figure can be viewed in the online issue which is available at wileyonlinelibrary.com]

Article Snippet: SOD1, Nluc and Rluc, were amplified from the SOD1 cDNA plasmid (Origene, Abingdon, UK, catalog no. SC111022), the pNL1.1 and pRL-null plasmid (Promega, Madison, WI), respectively.

Techniques: Expressing