Figure 1.

Adipogenesis is negatively regulated by AhR. A, the levels of the indicated proteins in mouse adipose tissues. B, protein levels of CUL4B and its related proteins were determined in 3T3-L1 cells. Upon differentiation with induction medium, cells were harvested and lysed for Western blotting. The protein band of interest is marked with a star. Signals from the Western blots were analyzed by Volume Analysis of Quantity One software with volume background subtraction. C, adipogenesis of AhR-overexpressing 3T3-L1 cells. Cells stably expressing empty vector or AhR were constructed using a lentivirus system. At day 9 postinduction, the cells were stained for lipid droplets using Oil Red O. D, adipogenesis of AhR-knockdown 3T3-L1. Stable cell lines were constructed using a lentivirus expressing shRNA (sh) for scrambled sequences (control) or mouse Ahr. On day 9 after induction of differentiation, cells were stained with Oil Red O. E, in AhR-overexpressing 3T3-L1 cells, total RNA samples were extracted at different time points upon induction. Samples were subjected to quantitative PCR analysis of adipogenic markers (CD36, Adipsin, and Fabp4). F, in AhR-knockdown 3T3-L1 cells, total RNA samples were extracted at different time points. Samples were subjected to quantitative PCR analysis of adipogenic markers (CD36, Adipsin, and Fabp4). Data are presented as mean ± S.D. (error bars); n = 4 with * representing p < 0.05, ** representing p < 0.01, and *** representing p < 0.001 by Student's t test. Ing-WAT, inguinal white adipose tissue; Epi-WAT, epididymal white adipose tissue; BAT, brown adipose tissue.