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: 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: