Exercise and Leptin Action Figure 6. Blockade of leptin induced inhibition of food intake by anti–IL-6 antibody. Hypothalami from rats were prepared as described in Research Design and Methods. (a) Tissue extracts from control and exercised rats were immunoblotted with anti–IL-6 antibody. (b) Leptin was injected intracerebroventricularly in control rats, exercised rats and exercised rats pretreated with anti–IL-6 at the doses indicated, and the animals were immediately exposed to food for a 12-hour period (n = 10–12 animals per group). (c, d, e, f) Representative Western blots of four independent experiments showing hypothalamic lysates from Wistar rats. Phospho-AMPKthr172, threonine-phosphorylated AMPK and total AMPK (c); phospho- ACCser79, serine phosphorylated ACC and total ACC (d); phospho-p70S6Kinasethr389, threonine phosphorylated p70S6Kinase and total p70S6K (e); phospho-4EBP1thr70, threonine phosphorylated 4EBP1 and total 4EBP1 (f). Data are the means6SEM. *p,0.05, vs. control, **p,0.05, vs. control plus leptin, #p,0.05, vs. exercise plus leptin. doi:10.1371/journal.pone.0003856.g006 PLoS ONE | www.plosone.org 10 December 2008 | Volume 3 | Issue 12 | e3856