Journal: Viruses

Article Title: Galectin-3-ITGB1 Signaling Mediates Interleukin 10 Production of Hepatic Conventional Natural Killer Cells in Hepatitis B Virus Transgenic Mice and Correlates with Hepatocellular Carcinoma Progression in Patients

doi: 10.3390/v16050737

Figure Lengend Snippet: Galectin-3 induced IL-10 production in hepatic cNK cells and prevented hepatocyte damage in HBs-Tg mice. MNCs were isolated from livers of 4- to 5-month-old HBs-Tg mice and control WT B6 mice. ( A ) IL-10 expression in hepatic cNK cells stimulated by rmGal-3. Liver MNCs (5 × 10 5 ) were stimulated by recombinant mouse Gal-3 (2.5 μg/mL or 5 μg/mL) in 200 μL of 10% FBS 1640 medium for 48 h. Cultured MNCs were harvested and analyzed by flow cytometry. cNK cells (CD3 − NK1.1 + CD49b + CD49a − ) were gated to analyze IL-10 expression. Histograms and MFI are shown. ( B ) Levels of IL-10 protein in culture supernatant. Liver MNCs (5 × 10 5 ) were stimulated by recombinant mouse Gal-3 (50 ng/mL) in 400 μL of 10% FBS 1640 medium for 48 h. Culture supernatant was collected and detected by IL-10 CBA flex set. ( C ) Monensin (2.5 μg/mL) was used to block secretion of IL-10 by cultured NK cells in vitro. Paired Student’s t test was used. * p < 0.05, ** p < 0.01. TD139 (Gal-3-INH) (15 μg/g body weight, once every 24 h for three times) was used. ( D ) Serum levels of IL-10 in HBs-Tg mice after TD139 (Gal-3-INH) treatment detected by ELISA. ( E ) IL-10 expression in liver tissues detected by IHC. Dark-brown dots represent positive staining. Bar = 50 μm. ( F ) Hepatic cNK cells (CD3 − NK1.1 + CD49b + CD49a − ) were gated and analyzed for expression of NKG2D. ( G ) Serum levels of ALT after TD139 (Gal-3-INH) treatment. Data are shown as mean ± SEM. Unpaired Student’s t test was used. * p < 0.05, ** p < 0.01.

Article Snippet: TD139 (TargetMol, Wellesley Hills, MA, USA) was dissolved in dimethyl sulfoxide (DMSO) (Solarbio, Beijing, China) and used for the inhibition of Galectin-3 in vivo (15 μg/g body weight, once per 24 h for three times).

Techniques: Isolation, Control, Expressing, Recombinant, Cell Culture, Flow Cytometry, Blocking Assay, In Vitro, Enzyme-linked Immunosorbent Assay, Staining

Journal: Neuroimmunomodulation

Article Title: Inhibition of Galectin-3 in a Rat Model of Epilepsy and Kainate-Activated BV2 Cells Limits Microglial Activation Through the NLRP3/Pyroptosis Pathway.

doi: 10.1159/000534833

Figure Lengend Snippet: Fig. 4. Inhibition of galectin-3 represses microglial activation in KA-treated microglia. a–f BV2 cells or primary microglia were treated with 100 μM KA for 24 h. For inhibition of galectin-3, BV2 cells or primary microglia were transfected with lentivirus vectors carrying Lgals3 shRNA or treated with 20 μM TD139 for 24 h. a–c The Iba1 expression was analyzed by immunofluorescence. d–f The protein expression of galectin-3 and Iba1 was analyzed by Western blot (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The EP model was successfully established when the seizure activity reached ≥ grade IV and continued for >30 min. For the EP + TD139 group, EP rats were injected intraperitoneally with TD139 (purity >98%; dissolve in dimethylsulfoxide; S0471, Selleck) at 8 mg/kg/d for 10 days starting 4 h after a seizure.

Techniques: Inhibition, Activation Assay, Transfection, shRNA, Expressing, Western Blot

Journal: Neuroimmunomodulation

Article Title: Inhibition of Galectin-3 in a Rat Model of Epilepsy and Kainate-Activated BV2 Cells Limits Microglial Activation Through the NLRP3/Pyroptosis Pathway.

doi: 10.1159/000534833

Figure Lengend Snippet: Fig. 5. Inhibition of galectin-3 represses the activation of NLRP3 inflammasome in KA-treated microglia. a–c BV2 cells or primary microglia were treated with 100 μM KA for 24 h. For inhibition of galectin-3, BV2 cells or primary microglia were transfected with lentivirus vectors carrying Lgals3 shRNA or treated with 20 μM TD139 for 24 h. The expression of NLRP3, ASC, C-caspase-1, and GSDMD-N was analyzed by Western blot (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The EP model was successfully established when the seizure activity reached ≥ grade IV and continued for >30 min. For the EP + TD139 group, EP rats were injected intraperitoneally with TD139 (purity >98%; dissolve in dimethylsulfoxide; S0471, Selleck) at 8 mg/kg/d for 10 days starting 4 h after a seizure.

Techniques: Inhibition, Activation Assay, Transfection, shRNA, Expressing, Western Blot

Journal: Frontiers in Chemistry

Article Title: The Structural Biology of Galectin-Ligand Recognition: Current Advances in Modeling Tools, Protein Engineering, and Inhibitor Design

doi: 10.3389/fchem.2019.00823

Figure Lengend Snippet: Galectins in complex with carbohydrate-derived monovalent inhibitors. (A) Thiodigalactoside “TD139” superimposed complexes of Gal-3 (magenta, PDB id: 5h9p) and Gal-7 (green, PDB id: 5h9q) (B) Taloside inhibitors superimposed complexes. Gal-1 and methyl 2-O-acetyl-3-O-toluoyl-beta-D-talopyranoside (cyan, PDB id: 3T2T), Gal-3 and methyl 3-deoxy-2-O-toluoyl-3-N-toluoyl-beta-D-talopyranoside (magenta, PDB id: 3T1M).

Article Snippet: Among these compounds, we can cite the C2-symmetric “TD139” (by Galecto Biotech) or its asymmetrical derivative “TAZTDG” by Hsieh et al. ( ).

Techniques: Derivative 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 + TD139 , 8 , SCI + TD139 (GAL3 inhibitor, 400 mg/kg, No. T899651g, Macklin).

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

Journal: Science Advances

Article Title: Galectin-3 exacerbates autoimmune diabetes by limiting regulatory T cell differentiation and function

doi: 10.1126/sciadv.adz7916

Figure Lengend Snippet: ( A ) Schematic diagram showing the protocol of TD139 treatment. ( B ) Incidence of diabetes expressed as percentage of diabetic mice at different ages ( n = 20). ( C ) Glucose excursion curve after receiving IPGTT ( n = 8). ( D ) Serum insulin concentration at 0 and 15 min during the IPGTT in 12-week-old TD139 or vehicle-treated NOD mice ( n = 7 to 8). ( E ) Representative H&E staining of pancreas from 12-week-old NOD mice treated with TD139 or vehicle ( n = 4). ( F ) Insulitis scores calculated on the basis of histological evaluation of pancreatic section as in (E). ( G ) Frequencies of CD4 + and CD8 + T cells in total CD3 + T cells from islets of 12-week-old NOD mice treated with TD139 or vehicle ( n = 4). ( H ) Frequencies of T reg cells (CD4 + CD25 + FOXP3 + ) from islets of 12-week-old NOD mice treated with TD139 or vehicle ( n = 4). ( I to K ) Frequencies of perforin + CD8 + T, Tc1 (CD8 + IFN-γ + ), Tc2 (CD8 + IL-4 + ), Tc17 (CD8 + IL-17 + ), and granzyme B + CD8 + T subsets from islets of 12-week-old NOD mice treated with TD139 or vehicle ( n = 3 to 4). ( L ) Frequencies of T H 1 (CD4 + IFN-γ + ), T H 2 (CD4 + IL-4 + ), and T H 17 (CD4 + IL-17 + ) from islets of 12-week-old NOD mice treated with TD139 or vehicle ( n = 4). Data are expressed as mean ± SEM. *P < 0.05 and **P < 0.01.

Article Snippet: TD139 (also known as GB0139), a small-molecule inhibitor of Galectin-3 in phase 1/2 clinical trials for idiopathic pulmonary fibrosis (IPF) ( NCT02257177 ) and COVID-19 pneumonitis ( NCT04473053 ), exhibits high affinity for the carbohydrate recognition domain of Galectin-3, preferentially targeting extracellular Galectin-3, due to its limited cell permeability ( – ).

Techniques: Concentration Assay, Staining