Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Spinal cord injury (SCI) increases galectin-3 (GAL3) expression in spinal neurons. (A) Relative GAL3 mRNA expression in the spinal cord after SCI. One-way ANOVA, n = 3/group. (B) Western blot analysis of GAL3 protein after SCI. (C) Statistical data show relative GAL3 protein expression after SCI. One-way ANOVA, n = 3/group. (D) Enzyme-linked immunosorbent assay (ELISA) detection of GAL3 protein levels in rat serum after SCI. One-way ANOVA, n = 6/group. (E) Immunofluorescence microscopy reveals GAL3 co-localization with NeuN post-SCI. (F) Fluorescence intensity of GAL3 after SCI. One-way ANOVA, n = 3/group. (G,H) Immunofluorescence double staining of GAL3 and GFAP (G) or IBA1 (H) after SCI. (I) Determination of optimal glutamate concentration and duration using CCK8 assay. (J) Relative GAL3 mRNA expression in the glutamate-stimulated spinal cord neurons. Unpaired Student’s t -test, n = 3/group. (K) Western blot analysis of GAL3 protein in neuronal injury model. (L) Relative GAL3 protein expression in neuronal injury model. Unpaired Student’s t -test, n = 3/group. (M) ELISA detection of GAL3 in cell supernatant of neuronal injury model. Unpaired Student’s t -test, n = 3/group. (N) Immunofluorescence microscopy showing GAL3 expression in neuronal injury model. (O) Quantification of GAL3 fluorescence intensity in neuronal injury model. Unpaired Student’s t -test, n = 3/group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibody used targeted the following proteins: NeuN (1:500, mouse IgG; No.26975, Proteintech), GAL3 (1:200, mouse IgG; No. 60207, Proteintech), and CDC42 (1:200, rabbit IgG; No.10155, Proteintech).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Microscopy, Fluorescence, Double Staining, Concentration Assay, CCK-8 Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) contributes to spinal cord injury (SCI)-induced motor impairment. (A) The mRNA level of GAL3 after siR-GAL3 treatment. Unpaired Student’s t -test, n = 3/group. (B) Western blot shows the protein level of GAL3 after siR-GAL3 treatment. (C) Statistical data show the knockdown of GAL3 by siRNA. Unpaired Student’s t -test, n = 3/group. (D) Enzyme-linked immunosorbent assay (ELISA) shows the secretory GAL3 in the supernatant of neurons after siRNA treatment. Unpaired Student’s t -test, n = 3/group. (E) The Basso-Beattie-Bresnahan (BBB) locomotor scores were increased after siR-GAL3 or inhibitor treatment. Two-way Repeated Measures ANOVA, n = 8/group. (F) The inclined plane angles were increased after siR-GAL3 or inhibitor treatment. Two-way Repeated Measures ANOVA, n = 8/group. When SCI + siR-GAL3 group was compared with SCI + Vehicle group, ** P < 0.01, *** P < 0.001; when SCI + TD139 group was compared with SCI + Vehicle group, # P < 0.05, ### P < 0.001; when SCI + GAL3 group was compared with SCI + Vehicle group, + P < 0.05,++ P < 0.01.

Article Snippet: The primary antibody used targeted the following proteins: NeuN (1:500, mouse IgG; No.26975, Proteintech), GAL3 (1:200, mouse IgG; No. 60207, Proteintech), and CDC42 (1:200, rabbit IgG; No.10155, Proteintech).

Techniques: Western Blot, Knockdown, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) is closely related to programmed cell death after spinal cord injury (SCI). (A) The four datasets before the batch effect were removed. (B) The four datasets after the batch effect were removed. (C) Volcano map shows DEGs in the SCI dataset. (D) Biological Process (BP) analysis of Gene Set Enrichment Analysis (GSEA) in the SCI dataset. Each column represents the P -value score of the pathway between the Sham group and the SCI group, with red indicating upregulation of the pathway in the SCI group, and blue indicating downregulation. (E) Protein-Protein Interaction Networks (PPI) analysis of differentially expressed genes (DEGs) in SCI dataset. In the PPI nodes, red signifies an increase in expression level, while blue indicates a decrease. The intensity of the color corresponds to the magnitude of the differential expression, with darker shades representing a higher differential expression multiple.

Article Snippet: The primary antibody used targeted the following proteins: NeuN (1:500, mouse IgG; No.26975, Proteintech), GAL3 (1:200, mouse IgG; No. 60207, Proteintech), and CDC42 (1:200, rabbit IgG; No.10155, Proteintech).

Techniques: Expressing, Quantitative Proteomics

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) regulates neuronal autophagy. (A) Western blot analysis of GAL3 and neuronal autophagy markers ATG7, P62, and LC3 II/I in neurons. (B-E) Quantification of western blot detection of GAL3 (B) , ATG7 (C) , P62 (D) , and LC3 II/I (E) in neurons. One-way ANOVA, n = 3/group. (F) Western blot analysis of GAL3 and neuronal autophagy markers ATG7, P62, and LC3 II/I in the spinal cord of rats. (G–I) Quantification of western blot detection of ATG7 (G) , P62 (H) , and LC3 II/I (I) in the spinal cord of rats. One-way ANOVA, n = 3/group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibody used targeted the following proteins: NeuN (1:500, mouse IgG; No.26975, Proteintech), GAL3 (1:200, mouse IgG; No. 60207, Proteintech), and CDC42 (1:200, rabbit IgG; No.10155, Proteintech).

Techniques: Western Blot

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Sequencing analysis of spinal cord neurons with galectin-3 (GAL3) knocked down. (A) Volcano map shows differential expression genes (DEGs) in the neuron dataset. (B) Biological process (BP) analysis of Gene Set Enrichment Analysis (GSEA) in the neuron dataset. Each column represents the P -value score of the pathway between the Sham group and the spinal cord injury (SCI) group, with red indicating upregulation of the pathway in the SCI group, and blue indicating downregulation. (C) The Protein-Protein Interaction Networks (PPI) analysis of DEGs in the neuron dataset. In the PPI nodes, red indicates that the expression level increases and blue indicates that the expression level decreases. The darker the color, the greater the differential expression multiple.

Article Snippet: The primary antibody used targeted the following proteins: NeuN (1:500, mouse IgG; No.26975, Proteintech), GAL3 (1:200, mouse IgG; No. 60207, Proteintech), and CDC42 (1:200, rabbit IgG; No.10155, Proteintech).

Techniques: Sequencing, Quantitative Proteomics, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) interacts with Cell-division-cycle-42 (CDC42) to regulate neuronal autophagy. (A) Intersected 29 core nodes from the neuron dataset with 22 core nodes from the spinal cord injury (SCI) dataset by the Venn diagram. (B) Correlation analysis between GAL3 and CDC42 expression level in SCI dataset. (C) Correlation analysis between GAL3 and CDC42 expression level in the neuron dataset. (D) Co-immunoprecipitation (Co-IP) shows a direct interaction between GAL3 and CDC42 in the glutamate-induced neuronal damage model. (E) Western blot shows the expression of CDC42, ATG7, P62, and LC3 II/I. (F–I) Quantification of western blot detection of CDC42 (F) , ATG7 (G) , P62 (H) , and LC3 II/I (I) . One-way ANOVA, n = 3/group. (J) Enzyme-linked immunosorbent assay (ELISA) detection of CDC42 in cell supernatant of GAL3-injury model. Unpaired Student’s t -test, n = 3/group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The primary antibody used targeted the following proteins: NeuN (1:500, mouse IgG; No.26975, Proteintech), GAL3 (1:200, mouse IgG; No. 60207, Proteintech), and CDC42 (1:200, rabbit IgG; No.10155, Proteintech).

Techniques: Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Cell-division-cycle-42 (CDC42) contributes to spinal cord injury (SCI)-induced motor function impairment. (A) The mRNA level after siR-CDC42 treatment. Unpaired Student’s t -test, n = 3/group. (B) Enzyme-linked immunosorbent assay (ELISA) shows the secretory CDC42 in the supernatant of neurons after siRNA treatment. Unpaired Student’s t -test, n = 3/group. (C) The Basso-Beattie-Bresnahan (BBB) locomotor scores were increased after siR-CDC42 and ML141 treatment. Two-way Repeated Measures ANOVA, n = 8/group. (D) The inclined plane angles were increased after siR-CDC42 and ML141 treatment. Two-way Repeated Measures ANOVA, n = 8/group. When SCI + siR-CDC42 group was compared with SCI + Vehicle group, * P < 0.05, ** P < 0.01, *** P < 0.001; when SCI + ML141 group was compared with SCI + Vehicle group, ## P < 0.01, ### P < 0.001. (E,F) Detection of the protein expression level of galectin-3 (GAL3) (E) and CDC42 (F) in serum of healthy volunteers and SCI patients by ELISA. Unpaired Student’s t -test, n = 8/group. *** P < 0.001.

Article Snippet: The primary antibody used targeted the following proteins: NeuN (1:500, mouse IgG; No.26975, Proteintech), GAL3 (1:200, mouse IgG; No. 60207, Proteintech), and CDC42 (1:200, rabbit IgG; No.10155, Proteintech).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Spinal cord injury (SCI) increases galectin-3 (GAL3) expression in spinal neurons. (A) Relative GAL3 mRNA expression in the spinal cord after SCI. One-way ANOVA, n = 3/group. (B) Western blot analysis of GAL3 protein after SCI. (C) Statistical data show relative GAL3 protein expression after SCI. One-way ANOVA, n = 3/group. (D) Enzyme-linked immunosorbent assay (ELISA) detection of GAL3 protein levels in rat serum after SCI. One-way ANOVA, n = 6/group. (E) Immunofluorescence microscopy reveals GAL3 co-localization with NeuN post-SCI. (F) Fluorescence intensity of GAL3 after SCI. One-way ANOVA, n = 3/group. (G,H) Immunofluorescence double staining of GAL3 and GFAP (G) or IBA1 (H) after SCI. (I) Determination of optimal glutamate concentration and duration using CCK8 assay. (J) Relative GAL3 mRNA expression in the glutamate-stimulated spinal cord neurons. Unpaired Student’s t -test, n = 3/group. (K) Western blot analysis of GAL3 protein in neuronal injury model. (L) Relative GAL3 protein expression in neuronal injury model. Unpaired Student’s t -test, n = 3/group. (M) ELISA detection of GAL3 in cell supernatant of neuronal injury model. Unpaired Student’s t -test, n = 3/group. (N) Immunofluorescence microscopy showing GAL3 expression in neuronal injury model. (O) Quantification of GAL3 fluorescence intensity in neuronal injury model. Unpaired Student’s t -test, n = 3/group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: SCI + GAL3 , 8 , SCI + GAL3 (400 mg/kg, No. HY- P77684 , MedChemExpress).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Microscopy, Fluorescence, Double Staining, Concentration Assay, CCK-8 Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) contributes to spinal cord injury (SCI)-induced motor impairment. (A) The mRNA level of GAL3 after siR-GAL3 treatment. Unpaired Student’s t -test, n = 3/group. (B) Western blot shows the protein level of GAL3 after siR-GAL3 treatment. (C) Statistical data show the knockdown of GAL3 by siRNA. Unpaired Student’s t -test, n = 3/group. (D) Enzyme-linked immunosorbent assay (ELISA) shows the secretory GAL3 in the supernatant of neurons after siRNA treatment. Unpaired Student’s t -test, n = 3/group. (E) The Basso-Beattie-Bresnahan (BBB) locomotor scores were increased after siR-GAL3 or inhibitor treatment. Two-way Repeated Measures ANOVA, n = 8/group. (F) The inclined plane angles were increased after siR-GAL3 or inhibitor treatment. Two-way Repeated Measures ANOVA, n = 8/group. When SCI + siR-GAL3 group was compared with SCI + Vehicle group, ** P < 0.01, *** P < 0.001; when SCI + TD139 group was compared with SCI + Vehicle group, # P < 0.05, ### P < 0.001; when SCI + GAL3 group was compared with SCI + Vehicle group, + P < 0.05,++ P < 0.01.

Article Snippet: SCI + GAL3 , 8 , SCI + GAL3 (400 mg/kg, No. HY- P77684 , MedChemExpress).

Techniques: Western Blot, Knockdown, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) is closely related to programmed cell death after spinal cord injury (SCI). (A) The four datasets before the batch effect were removed. (B) The four datasets after the batch effect were removed. (C) Volcano map shows DEGs in the SCI dataset. (D) Biological Process (BP) analysis of Gene Set Enrichment Analysis (GSEA) in the SCI dataset. Each column represents the P -value score of the pathway between the Sham group and the SCI group, with red indicating upregulation of the pathway in the SCI group, and blue indicating downregulation. (E) Protein-Protein Interaction Networks (PPI) analysis of differentially expressed genes (DEGs) in SCI dataset. In the PPI nodes, red signifies an increase in expression level, while blue indicates a decrease. The intensity of the color corresponds to the magnitude of the differential expression, with darker shades representing a higher differential expression multiple.

Article Snippet: SCI + GAL3 , 8 , SCI + GAL3 (400 mg/kg, No. HY- P77684 , MedChemExpress).

Techniques: Expressing, Quantitative Proteomics

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) regulates neuronal autophagy. (A) Western blot analysis of GAL3 and neuronal autophagy markers ATG7, P62, and LC3 II/I in neurons. (B-E) Quantification of western blot detection of GAL3 (B) , ATG7 (C) , P62 (D) , and LC3 II/I (E) in neurons. One-way ANOVA, n = 3/group. (F) Western blot analysis of GAL3 and neuronal autophagy markers ATG7, P62, and LC3 II/I in the spinal cord of rats. (G–I) Quantification of western blot detection of ATG7 (G) , P62 (H) , and LC3 II/I (I) in the spinal cord of rats. One-way ANOVA, n = 3/group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: SCI + GAL3 , 8 , SCI + GAL3 (400 mg/kg, No. HY- P77684 , MedChemExpress).

Techniques: Western Blot

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Sequencing analysis of spinal cord neurons with galectin-3 (GAL3) knocked down. (A) Volcano map shows differential expression genes (DEGs) in the neuron dataset. (B) Biological process (BP) analysis of Gene Set Enrichment Analysis (GSEA) in the neuron dataset. Each column represents the P -value score of the pathway between the Sham group and the spinal cord injury (SCI) group, with red indicating upregulation of the pathway in the SCI group, and blue indicating downregulation. (C) The Protein-Protein Interaction Networks (PPI) analysis of DEGs in the neuron dataset. In the PPI nodes, red indicates that the expression level increases and blue indicates that the expression level decreases. The darker the color, the greater the differential expression multiple.

Article Snippet: SCI + GAL3 , 8 , SCI + GAL3 (400 mg/kg, No. HY- P77684 , MedChemExpress).

Techniques: Sequencing, Quantitative Proteomics, Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Galectin-3 (GAL3) interacts with Cell-division-cycle-42 (CDC42) to regulate neuronal autophagy. (A) Intersected 29 core nodes from the neuron dataset with 22 core nodes from the spinal cord injury (SCI) dataset by the Venn diagram. (B) Correlation analysis between GAL3 and CDC42 expression level in SCI dataset. (C) Correlation analysis between GAL3 and CDC42 expression level in the neuron dataset. (D) Co-immunoprecipitation (Co-IP) shows a direct interaction between GAL3 and CDC42 in the glutamate-induced neuronal damage model. (E) Western blot shows the expression of CDC42, ATG7, P62, and LC3 II/I. (F–I) Quantification of western blot detection of CDC42 (F) , ATG7 (G) , P62 (H) , and LC3 II/I (I) . One-way ANOVA, n = 3/group. (J) Enzyme-linked immunosorbent assay (ELISA) detection of CDC42 in cell supernatant of GAL3-injury model. Unpaired Student’s t -test, n = 3/group. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: SCI + GAL3 , 8 , SCI + GAL3 (400 mg/kg, No. HY- P77684 , MedChemExpress).

Techniques: Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Galactin-3 regulation of CDC42 promotes neuronal autophagy following spinal cord injury

doi: 10.3389/fncel.2025.1622825

Figure Lengend Snippet: Cell-division-cycle-42 (CDC42) contributes to spinal cord injury (SCI)-induced motor function impairment. (A) The mRNA level after siR-CDC42 treatment. Unpaired Student’s t -test, n = 3/group. (B) Enzyme-linked immunosorbent assay (ELISA) shows the secretory CDC42 in the supernatant of neurons after siRNA treatment. Unpaired Student’s t -test, n = 3/group. (C) The Basso-Beattie-Bresnahan (BBB) locomotor scores were increased after siR-CDC42 and ML141 treatment. Two-way Repeated Measures ANOVA, n = 8/group. (D) The inclined plane angles were increased after siR-CDC42 and ML141 treatment. Two-way Repeated Measures ANOVA, n = 8/group. When SCI + siR-CDC42 group was compared with SCI + Vehicle group, * P < 0.05, ** P < 0.01, *** P < 0.001; when SCI + ML141 group was compared with SCI + Vehicle group, ## P < 0.01, ### P < 0.001. (E,F) Detection of the protein expression level of galectin-3 (GAL3) (E) and CDC42 (F) in serum of healthy volunteers and SCI patients by ELISA. Unpaired Student’s t -test, n = 8/group. *** P < 0.001.

Article Snippet: SCI + GAL3 , 8 , SCI + GAL3 (400 mg/kg, No. HY- P77684 , MedChemExpress).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing