Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition reduced lung tissues damage induced by lung I/R (A) The Gal3 protein expression in lung tissue was assessed by western blotting, and the density of Gal3 (relative to β-actin) on western blotting was quantified (n = 3). (B–D) The expression of Gal3 in lung tissue, BALF and serum was detected by ELISA (n = 6). (E) Paraffin-embedded lung tissue sections were stained with hematoxylin and eosin (n = 3) at magnifications of 200×, scale bar 200 μm and 400X, scale bar 100 μm. (F) The extent of lung injury was assessed by scoring the F images (n = 6 section per group). (G) The W/D ratio of lung tissue (n = 6). (H) Ultrastructural changes were evaluated through transmission electron microscopy. Arrows indicate lamellar body and triangles mitochondria (n = 3). Scale bar, 1 μm. The data were mean ± SEM. One-way analysis of variance was used for data comparison, ns, not significant, ∗p < 0.05, was statistically significant.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Transmission Assay, Electron Microscopy, Comparison

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition attenuated the inflammatory response induced by lung I/R (A–C) BALF of left lung after lung I/R was collected, and the supernatant was extracted to detect the contents of TNF-α, IL-1β, and IL-10 in BALF. (D–F) After LIRI, the heart blood was taken, and the supernatant was collected by centrifugation after standing at low temperature for 3 h. The levels of TNF-α, IL-1β, and IL-10 in the serum were detected. (G–I) The lung tissue was subjected to standard treatment to detect the contents of TNF-α, IL-1β, and IL-10 in lung tissue. Data shown are mean ± SEM from n = 6 per group, ns, no statistical significance, ∗p < 0.05, was statistically significant.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Centrifugation

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Inhibition of Gal3 reduces the expression of M1-type macrophages induced by lung I/R (A) Western blot of IL-1β, IL-6, TNF-α, and CCL5 in lung tissues. (B–E) Density analysis of IL-1β, IL-6, TNF-α, and CCL5 relative to β-actin in part A. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Lung tissues were stained with macrophages marker (F4/80, green), anti-IL-1β (red), and anti-IL-6 antibodies (red). Nuclei were stained with DAPI (blue). Magnification, 400×, scale bar, 100 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Expressing, Western Blot, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Inhibition of Gal3 increases the expression of M2 type macrophages induced by lung I/R (A) Western blot of Arg1, IL-10, Retnla, and Chil3 in lung tissues. (B–E) Density analysis of Arg1, IL-10, Retnla, and Chil3 relative to β-actin in part A. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Lung tissues were stained with macrophages marker (F4/80, green), anti-Arg1 (red), and anti-IL-10 antibodies (red). Nuclei were stained with DAPI (blue). Magnification, 400×, scale bar, 100 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Expressing, Western Blot, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition reduces macrophage polarization toward M1-type in OGD/R vitro model (A) Western blot of Gal3 in lung tissues. (B) Density analysis of Gal3 relative to β-actin in part A. (C) Cell viability was measured by CCK-8 assay (n = 6). (D) Western blot of IL-1β, IL-6, TNF-α, and CCL5 in RAW264.7 cells. (E–H) Densitometry of western blots in part D. Levels were standardized uniformly with β-actin. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (I and J) Immunofluorescence staining showed macrophages marker (F4/80, green), IL-1β (red), and IL-6 (red) in RAW264.7 cells. Nuclei were stained with DAPI (blue). Magnification, 200×, scale bar, 200 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Western Blot, CCK-8 Assay, Immunofluorescence, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition promotes the polarization of macrophages toward the M2-type in OGD/R vitro model (A) Western blot of Arg1, IL-10, Retnla, and Chil3 in RAW264.7 cells. (B–E) Densitometry of western blots in part A. Levels were standardized uniformly with β-actin. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Immunofluorescence staining showed macrophages marker (F4/80, green), Arg1 (red), and IL-10 (red) in RAW264.7 cells. Nuclei were stained with DAPI (blue). Magnification, 200×, scale bar, 200 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Western Blot, Immunofluorescence, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 and TREM2 colocalized in macrophages and Gal3 inhibited the expression of TREM2 in LIRI (A) Macrophage markers (F4/80, green), anti-TREM2 antibody (red), and anti-Gal3 antibody (pink) were used for immunofluorescence co-localization staining of lung tissue. Magnification, 400×, scale bar, 200 μm. (B) Enlarge the dotted box in part A. F4/80 (green), TREM2 (red), and Gal3 (pink) co-localization qualitative analysis. White dots to white triangles area (white dotted line) were analyzed with line intensity scans at higher magnification. Measurement of the intensity values demonstrated co-localization in those regions. Magnification, 2000×, scale bar, 50 μm. (C) Western blotting and (E) RT-qPCR expression of TREM2 in lung tissue. (D) Western blotting and (F) RT-qPCR expression of TREM2 in RAW264.7 cells. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05).

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Expressing, Immunofluorescence, Staining, Western Blot, Quantitative RT-PCR

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet:

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Recombinant, Modification, Saline, Protease Inhibitor, 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: 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

Journal: Scientific Reports

Article Title: Targeting fibroblast CD248 attenuates CCL17-expressing macrophages and tissue fibrosis

doi: 10.1038/s41598-020-73194-x

Figure Lengend Snippet: CD248 interacted with galectin-3 to induce CCL17-expressing pro-fibrotic macrophages. ( a ) qPCR of genes encoding cytokines and chemokines in macrophages isolated from D7 UUO kidneys of WT and Lgals3 knockout ( Lgals3 –/– ) mice. n = 5. ( b,c ) HEK293T cells were transfected with and without plasmid DNA expressing galectin-3 or CD248-DDK separately. CD248-DDK (DDK-IP) or galectin-3 (Gal3-IP) was immunoprecipitated from the cell lysates, and then immunoblot analyses of galectin-3 and CD248-DDK were performed. ( d ) Flow cytometry of the binding of CD248 extracellular domain (CD248ECD-EGFP) to WT and Lgals3 –/– BMDMs. Blue and red lines indicate results for WT and Lgals3 –/– BMDMs, respectively, with CD248ECD-EGFP. Black and gray lines indicate results for WT and Lgals3 –/– BMDMs, respectively, with control medium. ( e ) Flow cytometry of CD248ECD-EGFP binding to WT BMDMs in the presence of 25 mM lactose (red) or sucrose (blue). Conditioned medium from HEK293T cells in the presence of lactose (gray) or sucrose (black) was used as control. ( f ) qPCR of Ccl17 in WT and Lgals3 –/– BMDMs in the absence (control) or presence of 200 ng/ml rCD248 for 48 h. n = 6. ( g ) Gel plots of PCR for Ccr4 and Gapdh in D7 UUO-kidney myofibroblasts isolated from WT and Cd248 –/– mice. The reaction cycles for Ccr4 and Gapdh were 30 and 15, respectively. ( h ) qPCR for genes Col1a1 and Acta2 in isolated D7 UUO-kidney myofibroblasts after incubation with vehicle (Con), TGF-β1, and CCL17 for 24 h. Data are expressed as means ± standard errors of the mean. * P < 0.05, ** P < 0.01 and *** P < 0.001 by unpaired t-test in ( a ) and one-way ANOVA with post hoc Tukey’s multiple comparisons test in ( f , h ).

Article Snippet: HEK293T cells were transfected with plasmid pCMV6- Cd248 -DDK (MR210537; OriGene Technologies, Inc., Rockville, MD) or pCMV6- Lgals3 (MC208879; OriGene Technologies, Inc.) by PolyJet reagent (SignaGen Laboratories, Rockville, MD).

Techniques: Expressing, Isolation, Knock-Out, Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Flow Cytometry, Binding Assay, Incubation