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Image Search Results
Journal: Diabetes
Article Title: Gene Silencing of Phogrin Unveils Its Essential Role in Glucose-Responsive Pancreatic β-Cell Growth
doi: 10.2337/db08-0970
Figure Lengend Snippet: Effects of shRNA-expressing adenoviruses on GSIS and the proliferation of MIN6 cells. A : MIN6 cells were infected with adenoviruses integrating shVector, shPh1, shPh2, shPh3, or shIA2 for 72 h. The expression levels of endogenous phogrin and IA-2 proteins were determined by immunoblotting with specific antibodies for phogrin and IA-2. α-Tubulin is a nontargeting control. B : MIN6 cells infected with Ad-shVec, shPh1, shPh2, or shPh3 were immunostained with anti-phogrin antibody. C : MIN6 cells expressing shPh3 were fixed at time points 24, 48, and 72 h. Cells were then immunostained with anti-phogrin and anti-insulin antibodies. D : The infected cells were incubated for 2 h in modified KRB (2 mmol/l glucose) before high-glucose stimulation. After stimulation for 30 min with 2 or 25 mmol/l glucose, culture media ( left ; secretion) and cells ( right ; content) were collected and subjected to RIA for insulin. Data are given as means ± SE of five independent experiments. E : Time course of MIN6 cell proliferation 24–96 h after infection with Ad-shVec, shPh1, shPh2, shPh3, or shIA2. Cell number was measured as indicated in research design and methods . Phogrin-KD or IA2-KD cells grew more slowly than the shVec-infected cells (* P < 0.001, ** P < 0.005). F : Cells infected by Ad-shVec or shPh3 for 72 h were assayed for BrdU incorporation by immunofluorescence analysis. Experiments were performed four times ( n = 12). (Please see http://dx.doi.org/10.2337/db08-0970 for a high-quality digital representation of this figure.)
Article Snippet: Anti-adaptin, anti–carboxypeptidase E (anti-CPE), anti–calcium-dependent activator protein (anti-CAPS), anti-munc18, anti-dynamin, anti–clathrin heavy chain, anti–phosphatidylinositol 3-kinase (anti-PI3K), anti-Grb2, and anti–extracellular signal–related kinase (anti-ERK) mouse monoclonal antibodies and
Techniques: shRNA, Expressing, Infection, Western Blot, Incubation, Modification, BrdU Incorporation Assay, Immunofluorescence