Resistin in Dairy Cows: Plasma Concentrations during Early Lactation, Expression and Potential Role in Adipose Tissue
Figure 6
Immunolocalization and levels of resistin protein in bovine adipocytes.
Levels of mRNA (A) and protein (B) for resistin in isolated mature bovine adipocytes and in isolated bovine stromal vascular cells, as determined by qRT-PCR and western blotting, respectively. mRNA levels for CD68 (the main cell surface marker of macrophages) was also measured in in isolated mature bovine adipocytes and in isolated bovine stromal vascular cells (A). Relative expression was measured relatively to the geometric mean of 4 reference gene expression [cyclophilin A (PPIA), GAPDH, Actin B (ACTB), and ribosomal protein L19 (RPL19)] by real-time reverse-transcription PCR. Vinculin (VLC) was used as the loading control for immunoblotting. The results are representative of five cell preparations (adipose tissue from one cow was used for one cell preparation). Results are represented as means ± SEM. * indicates a significant difference (p<0.05). C. Subcutaneous white adipose tissue from 32-month-old cows was fixed and sectioned, and then subjected to immunochemical analysis to determine of the distribution of resistin. Bar: 100 μm. The image shown is representative of three experiments on three different animals.