Work flow of establishing obese mice model and proteomic analysis of BAT proteins.

<p>Female C57BL/6J mice were fed normal diet (ND) or high-fat diet (HFD) start at 6–8 weeks of age. Glucose tolerance test (OGTT) and hyperinsulinemic-euglycemic clamp were applied to assess the insulin sensitivity in mice after 22 weeks. The levels of blood lipid were tested as the auxiliary evaluation indexes of metabolic state. Proteins of BAT in ND and HFD group were quantified via iTRAQ-coupled 2D LC-MS/MS. After comparing the differential expressed proteins between HFD group and ND group, functional analysis of those differential proteins of BAT indicated the important role of BAT in obesity. Finally, western blot analysis was performed to validate the variation of three key differential proteins.