Journal: PLoS ONE

Article Title: Excessive Food Intake, Obesity and Inflammation Process in Zucker fa/fa Rat Pancreatic Islets

doi: 10.1371/journal.pone.0022954

Figure Lengend Snippet: A) qPCR analysis of the expression of TNFα, IL-1ß, IL6 and their receptors TNFR1, TNFR2, IL-1R1, IL6R and IFNγRα. Total mRNA were extracted from isolated fa/+ and fa/fa rats islets and converted to cDNA using reverse transcriptase and oligo dT primers. Cytokine or receptor-specific primers were then used to amplify the corresponding cDNA. Each bar represents the average +/− S.E. of twelve fa/fa (black square) and twelve fa/+ extracts (grey square). Asterisk denotes a significant difference between fa/fa and fa/+ extracts. B) Immunocytochemical analysis of cytokines and receptors expression; anti-TNF-R1, -TNF-R2, -TNFα, -IL-6R, -IL6, -IL-1R1, -IL-1R2, -IL-1ß, -IFNγ-Rα, -IFNγ-Rß and –IFNγ antibodies were used in double immunostaining experiments with anti-insulin antibody on fa/fa and fa/+ ß-cells. Staining of cytokines and receptors (red) was detected using Texas red conjugate and insulin (green) was revealed using FITC conjugated antibody. Pictures show merged images of the double staining, from six fa/fa and six fa/+ rat islets.

Article Snippet: In the immunofluorescence studies we used anti-cytokines antibodies against IL-1ß, IL-6, TNFα, IFNγ, and anti-cytokine receptor antibodies against IL-1R1, IL-1R2, IL-6R, TNF-R1, IFN-Rα, IFN-Rß (Santa Cruz Biotechnology, Santa Cruz, CA); guinea pig anti-insulin antibody was from MP Biomedicals (MP Biomedicals, Irvine, CA).

Techniques: Expressing, Isolation, Reverse Transcription, Double Immunostaining, Staining, Double Staining