Erythropoietin (EPO) has shown beneficial effects in the regulation of obesity and metabolic syndrome; however, the detailed mechanism is still largely unknown. Here, we created mice with adipocyte-specific deletion of EPO receptor. These mice exhibited obesity and decreased glucose tolerance and insulin sensitivity, especially when fed a high-fat diet. Moreover, EPO increased oxidative metabolism, fatty acid oxidation, and key metabolic genes in adipocytes and in white adipose tissue from diet-induced obese wild-type mice. Increased metabolic activity by EPO is associated with induction of brown fat-like features in white adipocytes, as demonstrated by increases in brown fat gene expression, mitochondrial content, and uncoupled respiration. Peroxisome proliferator-activated receptor (PPAR)a was found to mediate EPO activity because a PPARa antagonist impaired EPO-mediated induction of brown fat-like gene expression and uncoupled respiration. PPARa also cooperates with Sirt1 activated by EPO through modulating the NAD+ level to regulate metabolic activity. PPARa targets, including PPARg coactivator 1a, uncoupling protein 1, and carnitine palmitoyltransferase 1a, were increased by EPO but impaired by Sirt1 knockdown. Sirt1 knockdown also attenuated adipose response to EPO. Collectively, EPO, as a novel regulator of adipose energy homeostasis via these metabolism coregulators, provides a potential therapeutic strategy to protect against obesity and metabolic disorders. © 2013 by the American Diabetes Association.