Journal: PLoS ONE

Article Title: Bioactivity of a modified human Glucagon-like peptide-1

doi: 10.1371/journal.pone.0171601

Figure Lengend Snippet: Mouse pancreatic β cell proliferation (A), RT-qPCR analysis of gene expression (B), and insulin secretion (C) . MIN6 cells were seeded into flat-bottomed 96-well microtiter plates at a density of 6.0×10 4 cells per well. ( A ) Cell proliferation assay: cells were treated with various concentrations (0.3, 3, 15 and 30 μM) of mGLP-1 or GLP-1 for 48h, 10 μL CCK8 was then added to each well and incubated for 1 to 4 hours to determine the optimal reading of OD 450 . The control was cell cultures treated with DPBS only. ( B ) Relative expression of genes related with proliferation and apoptosis in cells treated with mGLP-1, GLP-1, or DPBS as determined by RT-qPCR (n = 3). ( C ) Insulin secretion assay: Mouse pancreatic β cells were incubated with 15 μM GLP-1 or mGLP-1 in the presence of glucose (10 mM) for 30, 60, 90, or 120 min. The controls were cells incubated with just DPBS in the presence of 10mM glucose. Cell culture supernatants were collected and insulin concentration determined using a mouse insulin ELISA kit (JiNingShiYe Ltd. Shanghai, China). Data shown represent the mean ±SD (n = 6). Different letters or (*) indicate significant difference at p <0.05.

Article Snippet: Mouse insulinoma cells (MIN6) (iCell Bioscience, Inc. Shanghai, China) were cultured in a 25 cm 2 cell culture flasks and then seeded into 96-well microplates at a density of 6.0×10 4 cells per well for the cell proliferation assay.

Techniques: Quantitative RT-PCR, Gene Expression, Proliferation Assay, Incubation, Control, Expressing, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Bioactivity of a modified human Glucagon-like peptide-1

doi: 10.1371/journal.pone.0171601

Figure Lengend Snippet: Cell viability ratio (CRV) of MIN6 cells treated with different concentrations of GLP-1 or mGLP-1.

Article Snippet: Mouse insulinoma cells (MIN6) (iCell Bioscience, Inc. Shanghai, China) were cultured in a 25 cm 2 cell culture flasks and then seeded into 96-well microplates at a density of 6.0×10 4 cells per well for the cell proliferation assay.

Techniques:

Journal: PLoS ONE

Article Title: Lack of Wdr13 Gene in Mice Leads to Enhanced Pancreatic Beta Cell Proliferation, Hyperinsulinemia and Mild Obesity

doi: 10.1371/journal.pone.0038685

Figure Lengend Snippet: A) Transfection with Ad Wdr 13 and AdGFP viruses shows overexpression of WDR13 protein in MIN6 cells as visualized by immunoblotting using anti WDR13 antibody. Lower panel shows beta actin as loading control. Overexpression of WDR13 protein results in retardation in cell proliferation after 48 h of transfection with 100 MOI. B) Overexpression of WDR13 protein results in accumulation of p21, whereas Cyclin D1, Cyclin D2, Cyclin E1 and p27 levels remain unaffected. C) siRNA knockdown of WDR13 in MIN6 cells. MIN6 cells were transfected with nonspecific scrambled (Scr) siRNAs and WDR13 specific siRNA. Immunoblot analysis shows Knockdown of WDR13 protein. Actin was used as loading control. Immunoblot, using p21 antibody shows reduction of p21 levels in WDR13 knockdown MIN6 cell. D) Cell cycle inhibitor p21 expression in purified pancreatic islets of Wdr 13 knockout mice and that of wild type littermates by western blot analysis. Beta actin was used as loading control. p21 expression is less in the islets of knockout mice. E) Occupancy by WDR13 at p21 promoter revealed by chromatin immunoprecipitation using primers specific for p21 and GAPDH.

Article Snippet: 10,000 MIN6 cells (obtained from National Centre for Cell Science, Pune, India) were seeded per well of 24 well plates in DMEM media containing 10% FBS and were transfected either with AdGFP or Ad Wdr13 using a titer of 100 MOI.

Techniques: Transfection, Over Expression, Western Blot, Control, Knockdown, Expressing, Purification, Knock-Out, Chromatin Immunoprecipitation