Effect of CG on expression of adipogenesis-related genes. 3T3-L1 preadipocytes were differentiated into adipocytes in DMII medium with the absence or presence of 10 μg/ml or 100 μg/ml CG for 4 or 6 days. Con, 3T3-L1 preadipocytes; DMII, fully differentiated adipocytes (0.5 mM 3-isobutyl-1-methylxanthine, 100 μM indomethason, 0.25 μM dexamethasone and 167 nM insulin); 10 μg/ml, fully differentiated adipocytes (DMII + 10 μg/ml CG); 100 μg/ml, fully differentiated adipocytes (DMII + 100 μg/ml CG). (A) CG inhibited the expression of adipocyte-specific transcription factors during differentiation. The gene expression was performed by RT-PCR and all gene expression was normalized using β-actin as reference gene. All experiments were performed in three independent experiments. (*) p < 0.05, (**) P < 0.01, (***) P < 0.001 versus the control (DMII) group at each gene expression. (B) CG attenuated the expression of adipogenesis-related genes in 3T3-L1 adipocytes. Total cell lysates were isolated from 3T3-L1 adipocytes at day 4 or day 6 after induction of differentiation. Immunoblotting analysis was performed as described in Methods. (C) The relative expression of adipogenesis-related genes after treating CG for 4 or 6 days. All gene expressions were normalized using β-actin as a reference gene. The data shown are representative of three independent experiments. (*) p < 0.05, (**) P < 0.01, (***) P < 0.001 versus the control (DMII) group at each gene expression.