Endocrine disrupting chemical-induction of adipocyte protein expression. 3T3-L1 differentiation was induced by incubation of confluent preadipocytes for 3 days in 10% fetal bovine serum (FBS) in Dulbecco’s modified Eagle’s medium (DMEM) containing 100 nmol/l dehydrocorticosterone, 167 nmol/l insulin, and 0.5 mmol/l isobutylmethylxanthine alone (Diff) or supplemented with dexamethasone (Dex), bisphenol A (BPA), dicyclohexyl phthalate (DCHP), endrin, or tolylfluanid (TF) at concentrations ranging from left to right from 1 µmol/l to 100 pmol/l, with each well representing a progressive tenfold dilution. The cells were subsequently treated for an additional 2 days in 10% FBS in DMEM containing 167 nmol/l insulin. Whole-cell lysates were prepared, and the protein expression pattern was analyzed by immunoblotting for insulin receptor subunit α (IR-α), CCAAT/enhancer binding protein α (c/EBPα), adiponectin, and β-actin after resolution by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Protein expression was also compared to undifferentiated preadipocytes maintained in 10% FBS in DMEM (FBS). Results shown are representative of three independent experiments.