gal3 inhibitor  (TargetMol)

Journal: Nature Communications

Article Title: Galectin-3 impairs calcium transients and β-cell function

doi: 10.1038/s41467-024-47959-1

Figure Lengend Snippet: a Gal3 levels in plasma of NC and HFD-fed mice ( n = 6 mice). b Gal3 levels in plasma of db/m and db/db mice ( n = 3 mice, 7 to 14 weeks of age). c , d Confocal image of CD11c by immunostaining in insulin-positive cells in pancreatic sections from NC and HFD-fed mice ( c ) or from db/m and db/db mice ( d ). n = 6 islets in 3 pancreatic sections, Scale bar, 50 μm. e , f Inflammatory gene expression in pancreas from HFD-fed ( e ) and db/db ( f ) mice. n = 4 mice for Lgals3 and Adgre1 and n = 5 mice in e for other genes. n = 4 mice for db/m and n = 6 mice in ( f ) for db/db . g , h Gal3 levels in primary islets from HFD-fed ( g ) and db/db ( h ) mice. n = 10 mice ( g ), n = 4 mice ( h ). i GSIS in primary islets from NC and HFD-fed mice (n = 6 biologically independent samples). j GSIS in primary islets from db/m and db/db mice (n = 4 biologically independent samples). NC mice 20 weeks of age, HFD: 12 weeks from 8 weeks of age; db/m and db/db 28 weeks of age ( a , c – j ). Data were analyzed by two-sided Student’s t -test without adjustments for multiple comparisons. All data are presented as the mean ± SEM. Source data are provided as Source Data file. NC Normal chow.

Article Snippet: Mice were treated with the Gal3 inhibitor GB1107 (MCE, HY-114409, 10 mg/kg per day ) or vehicle (0.1% Tween 80 distilled water) by oral gavage.

Techniques: Immunostaining, Expressing

Journal: Nature Communications

Article Title: Galectin-3 impairs calcium transients and β-cell function

doi: 10.1038/s41467-024-47959-1

Figure Lengend Snippet: a – d Glucose-stimulated insulin secretion (GSIS) in INS-1 cells ( a ), MIN6 cells ( b ), NC mice primary islets ( c ), and db/db mice primary islets ( d ) with Gal3 treatment (MIN6 cells 80 ng/ml, INS-1 cells and islets 250 ng/ml) for 6 h. Gal3 existed all the time in the experimental process. (( a , c , d ), n = 4 biologically independent cell samples; ( b ), n = 6 biologically independent cell samples.) e Islet perfusion experiment in islets from 12-week-old NC mice with Gal3 (250 ng/ml) treatment for 6 h. f KCl (30 mM) stimulated insulin secretion in mice islets with or without Gal3 (250 ng/ml, 1 h) treatment ( n = 4 biologically independent cell samples). g Arginine (10 mM) stimulated insulin secretion in mouse islets with glucose (16.8 mM) and Gal3 (250 ng/ml, 1 h) treatment ( n = 5 biologically independent cell samples). (Islets from 12-week-old NC mice ( c , e , f , g ) and from 17-week-old db/db mice ( d )). h – i Confocal image of Gal3 colocalization with CD11c by immunostaining in insulin-positive cells in pancreatic sections from HFD-fed mice ( h ) or from db/db mice ( i ). n = 6 islets in 3 pancreatic sections (HFD: 16 weeks from 8 weeks of age, db/db 12 weeks of age). j Sketch map of MIN6 cells and MΦ coculture system and immunofluorescence staining of MIN6 cells and MΦ in the coculture system. Insulin (Green) labelled MIN6 cells and CD11c (Red) labelled MΦ. The MΦ were peritoneal macrophages from WT or Gal3 KO mice on a HFD feeding 12 weeks. n = 3 independent experiments. k Proinflammatory genes expression in MIN6 and MΦ from HFD-fed mice coculture system ( n = 6 for MIN6 group and n = 8 biologically independent cell samples for MIN6 + MΦ groups). l Gal3 levels in condition medium from coculture system with MΦ isolated from Gal3 KO or WT mice on a HFD. m GSIS in MIN6 and MΦ coculture system. n = 8 biologically independent cell samples ( l , m ). Data were analyzed by two-sided Student’s t -test without adjustments for multiple comparisons. All data are presented as the mean ± SEM. Source data are provided as Source Data file.

Article Snippet: Mice were treated with the Gal3 inhibitor GB1107 (MCE, HY-114409, 10 mg/kg per day ) or vehicle (0.1% Tween 80 distilled water) by oral gavage.

Techniques: Immunostaining, Immunofluorescence, Staining, Expressing, Isolation

Journal: Nature Communications

Article Title: Galectin-3 impairs calcium transients and β-cell function

doi: 10.1038/s41467-024-47959-1

Figure Lengend Snippet: a Blood Gal3 levels in WT, Gal3 +/− and Gal3 −/− mice on HFD-fed ( n = 8 mice). b – e Body weight ( b , n = 12 WT mice, n = 7 Gal3 +/− mice), intravenous glucose tolerance test (IVGTT) ( c ), Area under curve (AUC) during IVGTT ( d ) and first-phase insulin secretion ( e ) in WT and Gal3 +/− mice on HFD-fed after 6 h of fasting. c – e , n = 9 WT mice, n = 7 Gal3 +/− mice. f – i Plasma glucose ( f ), Insulin level ( g ), The AUC of the first phase of insulin secretion (from 0 to 10 min) and the second insulin secretion (from 10 to 120 min) ( h ), and GIR ( i ) in hyperglycemic clamp study in Gal3 +/− mice on HFD-fed. n = 6 mice. j GSIS in primary islets from Gal3 +/− mice on HFD-fed ( n = 6 biologically independent samples). HFD: 8–12 weeks from 8 weeks of age ( c – j ). k Experimental scheme of the db/db and db/db GKO mice. l , m Gal3 concentration in plasma ( l , n = 5 db/db mice, n = 6 db/db GKO mice) and islets ( m , n = 10 db/db mice, n = 5 db/db GKO mice). n – r ITT ( n ), IPGTT ( o ), Body weight ( p ), time course study of first-phase insulin secretion ( q ), and islets GSIS ( r ) in db/db and db/db GKO mice. n = 5 db/db mice, n = 7 db/db GKO mice ( n – q ); n = 8 ( db/db ), n = 5 ( db/db GKO) biologically independent samples ( r ). The age of mice ( l – r ) was same with ( k ). Data were analyzed by two-sided Student’s t -test without adjustments for multiple comparisons. All data are presented as the mean ± SEM. Source data are provided as Source Data file.

Article Snippet: Mice were treated with the Gal3 inhibitor GB1107 (MCE, HY-114409, 10 mg/kg per day ) or vehicle (0.1% Tween 80 distilled water) by oral gavage.

Techniques: Concentration Assay

Journal: Nature Communications

Article Title: Galectin-3 impairs calcium transients and β-cell function

doi: 10.1038/s41467-024-47959-1

Figure Lengend Snippet: a Experimental scheme of HFD mice by clodronate injection. b Gal3 level in blood. c – f IPGTT ( c ), AUC of IPGTT ( d ), first-phase insulin secretion ( e ) and GSIS in primary islet from clodronate- or control-treated HFD mice ( f ). n = 6 mice ( b – e ). g – j Gal3 levels in the blood ( g , n = 5 mice), IPGTT ( h ), first-phase insulin secretion ( i ), and GSIS in primary islets ( j ) in HFD-fed mice, HFD: 4 weeks from 8 weeks of age. n = 6 mice (Gal3 f/f and Gal3 f/+ - lyz cre), n = 7 mice (Gal3 f/f - lyz cre) ( h , i ), n = 6 biologically independent samples ( f , j ). k Immunohistochemistry analysis of insulin (green) and glucagon (red) in pancreas of WT and Gal3 +/− mice on HFD-fed. Scale bar, 20 μm. l β-cell area in ( k ). m β-cell mass in WT and Gal3 +/− on HFD-fed mice. n = 4 islets in 4 pancreatic sections (HFD: 16 weeks from 8 weeks of age ( k – m ). n Immunohistochemistry analysis of insulin (green) and glucagon (red) in pancreas of db/db and db/db GKO mice. Scale bar, 20 μm. o β-cell area of ( n ). p β-cell mass in db/db and db/db GKO mice. n = 8 islets in 5 pancreatic sections (16 weeks of age, n – p ). q – s mRNA level of Ins1 ( q ), MafA ( r ) and Nkx6.1 ( s ) in islets from WT and Gal3 +/− mice on a HFD feeding (16 weeks feeding from 8 weeks of age). mRNA level of each gene was normalized to the 36B4 mRNA level in the same sample. n = 4 mice (WT in ( q – s ) and Gal3 +/− in r ), n = 5 mice (Gal3 +/− ) ( q , s ). Mice information ( b – e , g – i ) was same with ( a ). Data were analyzed by two-sided Student’s t -test without adjustments for multiple comparisons. All data are presented as the mean ± SEM. Source data are provided as Source Data file. P -values in red in ( h ) represent Gal3 f/f - lyz Cre vs Gal3 f/f mice; P -values in orange in ( h ) represent Gal3 f/+ - lyz Cre vs Gal3 f/f mice.

Article Snippet: Mice were treated with the Gal3 inhibitor GB1107 (MCE, HY-114409, 10 mg/kg per day ) or vehicle (0.1% Tween 80 distilled water) by oral gavage.

Techniques: Injection, Control, Immunohistochemistry

Journal: Nature Communications

Article Title: Galectin-3 impairs calcium transients and β-cell function

doi: 10.1038/s41467-024-47959-1

Figure Lengend Snippet: a Typical traces of time-lapse calcium imaging via fluo-4, AM (1 μM) in MIN6 cells with high glucose (16.8 mM) stimulation and Gal3 (80 ng/ml, 6 h) treatment. n = 33 cells in vehicle group, and n = 26 cells in Gal3 group. b Frequency of cytosolic calcium spikes of ( a ). c – e Cytosolic calcium dynamics ( c ), frequency of cytosolic calcium spikes ( d ) and peak value of cytosolic calcium transient ( e ) stimulated by high glucose (16.8 mM) in different β cells of Ins1-GCaMP6f mouse islets with or without Gal3 (250 ng/ml, 6 h) treatment. Scale bar, 10 μm. n = 4 biologically independent samples (12-week-old NC-fed mice, c – e ). f, g The ratios of fluorescence signals ( f ) and distribution analysis of concentrations of cytosolic free Ca 2+ ([Ca 2+ ]i) ( g ) by fura-2, AM (2 μM) at 340 nm and 380 nm in MIN6 cells with basal glucose (2.8 mM) and Gal3 (80 ng/ml, 6 h) treatment. n = 222 cells in vehicle group, and n = 232 cells in Gal3 group. h Diagram of patch clamp experiment, created with BioRender.com. i , j Ca 2+ currents records in MIN6 cells without Gal3 and sequentially given Gal3 (80 ng/ml, 5 min) ( n = 4 biologically independent cell samples). k Ca 2+ currents records in MIN6 cells without Gal3 and sequentially given Gal3 (80 ng/ml, 5 min) and then given vehicle buffer washout. Data were analyzed by two-sided Student’s t -test without adjustments for multiple comparisons. All data are presented as the mean ± SEM. Source data are provided as Source Data file.

Article Snippet: Mice were treated with the Gal3 inhibitor GB1107 (MCE, HY-114409, 10 mg/kg per day ) or vehicle (0.1% Tween 80 distilled water) by oral gavage.

Techniques: Imaging, Fluorescence, Patch Clamp

Journal: Nature Communications

Article Title: Galectin-3 impairs calcium transients and β-cell function

doi: 10.1038/s41467-024-47959-1

Figure Lengend Snippet: a Co-immunoprecipitation of Gal3-GFP and Flag-tagged CACNG1 ( n = 3 independent experiments). b Colocalization of Gal3 and CACNG1 in islets from db/db (17 weeks of age) and HFD (16 weeks feeding from 8 weeks of age) mice, n = 3 islets in 3 pancreatic sections. c Co-immunoprecipitation of Gal3 and CACNG1 in MIN6 and macrophage coculture system. d Gal3 and CACNG1 were colocalization on the membrane in MIN6 cells. e Direct interaction of Gal3 and CACNG1 in the PLA. Scale bar, 5 μm. f The Gal3 and CACNG1 binding affinity assay was performed using a BLI system. g Gal3-tracing approach. Scale bar, 20 μm and 5 μm. h Co-immunoprecipitation of Gal3-GFP and Flag-tagged WT CACNG1, single amino acid mutant CACNG1 (N43A), CACNG1 (N80A), and double amino acid mutant CACNG1 (N43A/N80A). i Co-immunoprecipitation of Gal3-GFP and Flag-tagged CACNG1. n = 1 ( c ), n = 3 ( d , e , g – i ) independent experiments. j Effects of Gal3 (80 ng/ml, 30 min) and CBP (30 min) on high glucose (16.8 mM) stimulated calcium signals. k Peak value of cytosolic calcium transients in ( j ). n = 20 cells ( j – k ). l GSIS in islets with Gal3 (250 ng/ml, 1 h) and CBP (1 h) treatment. m GSIS in islets from WT and CKO mice with or without Gal3 (250 ng/ml, 1 h) treatment. n GSIS in Cacng1 f/f and Cacng1 Ins1Cre-GCaMP6f ( Cacng1 βKO ) mice islets with or without Gal3 (250 ng/ml, 1 h) treatment. n = 6 ( l ), n = 7 ( m ), n = 8 ( n ) biologically independent samples). Cytosolic calcium dynamics in β cells from Cacng1 βKO mice islets with or without Gal3 (250 ng/ml, 1 h) ( n = 25 cells). Islets are from 10-week-old NC mice ( l – o ). Data were analyzed by two-sided Student’s t -test without adjustments for multiple comparisons. All data are presented as the mean ± SEM. Source data are provided as Source Data file. CBP, CACNG1 blocking peptide (1 μg/ml).

Article Snippet: Mice were treated with the Gal3 inhibitor GB1107 (MCE, HY-114409, 10 mg/kg per day ) or vehicle (0.1% Tween 80 distilled water) by oral gavage.

Techniques: Immunoprecipitation, Membrane, Binding Assay, Mutagenesis, Blocking Assay

Journal: Nature Communications

Article Title: Galectin-3 impairs calcium transients and β-cell function

doi: 10.1038/s41467-024-47959-1

Figure Lengend Snippet: a – c Effect of Gal3 (80 ng/ml, 1 h) and Gal3 inhibitors GB1107 (1 μM, 1 h), TD139 (1 μM, 1 h) ( a ) and Cpd47 (1 μM, 1 h) ( b ) and Gal3 neutralizing antibody (Ab) (1 μM, 1 h) ( c ) on GSIS in MIN6 cells. n = 6 ( a ), n = 5 ( b ), n = 4 ( c ) biologically independent cell samples). d Effects of Gal3 (80 ng/ml, 30 min) and GB1107 (1 μM, 30 min) on high glucose (16.8 mM) stimulated calcium signals in MIN6 cells. e , Peak value of cytosolic calcium transient in MIN6 with or without Gal3 (80 ng/ml, 30 min) and GB1107 (1 μM, 30 min). n = 20 cells ( d , e ). f – l Experimental scheme ( f ), body weight ( g ), IPGTT ( h ), AUC of GTTs ( i ), ITT ( j ), in vivo GSIS ( k ), and GSIS of isolated primary islets ( l ) in db/db mice treated with GB1107 (10 mg/kg). n = 10 mice ( f – k ), n = 5 Vehicle and n = 6 GB1107 biologically independent samples ( l ). m Experimental scheme of db/db mice by Gal3 antibody injection. At the age of 5 weeks, db/db mice were intravenously injected with Gal3 antibodies every 3 days for a total of 5 consecutive injections. Additional injections were administered at the 13th and 14th weeks. n – q Body weight ( n , n = 10 mice), IPGTT (15 weeks of age) ( o , n = 9 mice), and GSIS in primary islets (16 weeks of age) ( p , n = 4 biologically independent samples) from db/db mice. q LGALS3 gene expression in primary islets from healthy persons and diabetic patients ( n = 7 biologically independent samples). r , s GSIS in human primary islets from healthy persons ( r ) and diabetic patients ( s ) with hGal3 (250 ng/ml, 6 h) treatment. t GSIS in islets from healthy persons after hGal3 (250 ng/ml, 6 h) and Gal3 inhibitor Cpd47 (1 μM, 6 h) treatment. n = 5 ( r – t ) biologically independent samples). Data were analyzed by two-sided Student’s t -test without adjustments for multiple comparisons. All data are presented as the mean ± SEM. Source data are provided as Source Data file. Mice information ( g – l ) was same with ( f ).

Article Snippet: Mice were treated with the Gal3 inhibitor GB1107 (MCE, HY-114409, 10 mg/kg per day ) or vehicle (0.1% Tween 80 distilled water) by oral gavage.

Techniques: In Vivo, Isolation, Injection, Expressing