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. 2013 Oct 18;62(11):3718–3726. doi: 10.2337/db13-0050

FIG. 5.

FIG. 5.

ATMs in epididymal WAT of PfnWT and PfnHet. A: Flow cytometry analysis of SVF (described in detail in research design and methods) included staining with CD45 (leukocytes), CD11b, and F4/80 (ATMs, referred to as Macs) after gating for propidium iodide-negative (live) cells. Subsequently, protein lysates were obtained from ATMs sorted under sterile conditions and analyzed by Western blot (inset). The expected 50% reduction in pfn was observed in ATMs from PfnHet. B: Quantification of the ATM/CD45+ ratio demonstrates a statistically significant decrease of ATMs in PfnHet (n = 5). *P < 0.001. C: CLSs (asterisks) were detected in sections from epididymal WAT after staining with anti-F4/80 antibody followed by colorimetric detection (brown). Sections were counterstained with hematoxylin (blue). The negative control is a consecutive HFD section stained with nonimmune IgG. Scale bar: 25 μm. D: CLS density (per 400 adipocytes) showing a statistically significant decrease in CLS (shown with * in C) in PfnHet-HFD compared with PfnWT-HFD (n = 5). *P < 0.001. E: Mean adipocyte size area was comparable in PfnWT- and PfnHet-HFD for 12 weeks (n = 5). F: Distribution of adipocyte by size category shows similar frequencies for PfnWT and PfnHet.