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Russell, C. , Petersen, R. , Rao, S. , Ricci, M. , Brolin, R. , and Fried, S. K. (1998) Leptin expression in adipose tissue from obese humans: depot-specific regulation by insulin and dexamethasone. Am. J. Physiol. 275, E507–E515. 67. Bouchard, C. (1997) Genetic determinants of regional fat pad distribution. Human Reprod. 12, 1–5. 68. , Revelli, J. , and Pénicaud, L. (1993) Adipose tissues from various anatomical sites are characterized by different patterns of gene expression and regulation. Biochem.

FASEB J. 4, 2890–2898. 14. Rothwell, N. J. and Stock, M. J. (1986) Brown adipose tissue and diet-induced thermogenesis, in Brown Adipose Tissue (Trayhurn, P. , D. , eds. , London, pp. 269–298. 15. Trayhurn, P. (1989) Thermogenesis and the energetics of pregnancy and lactation. Can. J. Physiol. Pharmacol. 67, 370–375. 16. , Flier, J. , and Lowell, B. B. (1998) Obesity after genetic ablation of brown adipose tissue. Z. Ernährungswiss 37, 1–7. 17. Nicholls, D. G. and Locke, R. M. (1984) Thermogenic mechanisms in brown fat.

1. METHOD 1 (SEE NOTES 4,5) 1. 4, 4 h at 4°C. 2. Wash overnight in PB. 3. 4, 1 h at 4°C. 4. Wash in 1X PBS. 2. METHOD 2: OSO4 MACERATION TECHNIQUE 1. Quick perfusion (when this is possible) with the fixative indicated at item 3. 2. Reduction of the tissue into thin strips (1 × 1 × 5 mm). 3. 1 M Na-cacodylate-HCl buffer for 15 min at RT in the dark. 4. 2. 5. 25% potassium ferrocyanide for 2 h (36). 6. Embedding in agarose and sectioning (about 150 µm) with a chopper microtome. 7. Washing in PBS.

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