w 5 hydrochloride  (Tocris)

Journal: Diabetologia

Article Title: Single-cell RNA sequencing identifies endothelial-derived HBEGF as promoting pancreatic beta cell proliferation in mice via the EGFR– Kmt5a –H4K20me pathway

doi: 10.1007/s00125-024-06341-y

Figure Lengend Snippet: Kmt5a –H4K20me is involved in the proliferation of beta cells through transcriptional regulation of Cdk1 . ( a ) Heatmap analysis of the expression of histone-modifying enzymes in beta cell subclusters isolated from mouse pancreatic islets, including histone methyltransferases and acetyltransferases. The x axis represents the subclusters of beta cells, and the y axis represents the different histone-modifying enzymes. ( b , c ) Violin plots of the distributions of the histone-modifying enzymes Kmt5a , Ezh2 and Hat1 in the beta cell subclusters ( b ), and gene expression levels in SC5 beta cells at the different time points ( c ). ( d ) Western blot analysis of H4K20me and MKI67 expression in islets from nulliparous and GD14.5 mice and associated quantification graph. β-actin was used as an internal control. ( e ) Immunofluorescence analysis of EdU incorporation (green) in MIN6 cells after treatment with 20 μmol/l UNC0379 or an equal volume of DMSO (control). Scale bar, 20 μm. ( f ) Number of EdU-positive MIN6 cells after treatment with UNC0379 at final concentrations of 0, 10, 15 and 20 μmol/l for 6 h. ( g ) CCK8 analysis of MIN6 cell viability after exposure to UNC0379 at final concentrations of 0, 10, 15 and 20 μmol/l for 6 h. ( h ) Western blot analysis of H4K20me and MKI67 expression in INS1 cells treated with UNC0379 at final concentrations of 0, 10, 15 and 20 μmol/l for 6 h. β-actin was used as an internal control. ( i , j ) Violin plots showing the distribution of Cdk1 in the beta cell subclusters ( i ) and Cdk1 expression in SC5 beta cells at the different time points ( j ). ( k ) Relative expression of Kmt5a and Cdk1 vs β-actin in MIN6 cells transfected with siRNA-control and siRNA- Kmt5a . ( l ) Chromatin immunoprecipitation analysis. The schematic shows the relative positions of the primers in the chromatin immunoprecipitation assays. A H4K20me-specific antibody was used in the analysis and primers were designed in the promoter regions of Cdk1 . Data are presented as mean ± SEM from three independent experiments. * p <0.05, ** p <0.01. TSS, transcription start site

Article Snippet: INS1 cells (CL-0368, Procell, China) were cultured in RPMI 1640 medium with 10% (vol./vol.)

Techniques: Expressing, Isolation, Gene Expression, Western Blot, Control, Immunofluorescence, Transfection, Chromatin Immunoprecipitation

Journal: Biomolecules

Article Title: PDX-1: A Promising Therapeutic Target to Reverse Diabetes

doi: 10.3390/biom12121785

Figure Lengend Snippet: Representative potential PDX-1 inducing drugs, small molecules and natural compounds.

Article Snippet: 1 , Swietenine (Stn) and swietenolide (Std) , INS-1 cells (Procell CL-0368) , 2 μM, 5 μM, 8 μM, 10 μM, 15 μM, 20 μM, 30 μM, 40 μM, 50 μM , It up-regulates the expression of PDX-1 protein, improves the insulin secretion function, protects oxidative stress injury, and reduces apoptosis. , [ ] .

Techniques: Expressing, Purification, Isolation, DNA Methylation Assay, Cell Differentiation, Phospho-proteomics, Gene Expression, Histone Deacetylase Assay, Methylation