Figure 2.

FTY720 phosphate (FTY-p) decreases lipid droplets during the differentiation of 3T3-L1 preadipocytes. (A) Preadipocytes were induced to differentiate with FTY-p of increasing concentrations for 6 days. The AdipoRed assays were performed on day 6 and were photographed with a light microscope (× 200). (B) Fluorescence was measured with an excitation wavelength of 485 nm and an emission wavelength of 572 nm. Bar graph indicates the mean ± S. E. M. (n = 3). The experiments were repeatedly performed to confirm the results. The data were analyzed using analysis of variance (ANOVA) and Duncan multiple range test (P < 0.05). Means sharing a common alphabetical symbol do not differ significantly. (C-E) Preadipocytes were induced to differentiate with FTY-p (1 µM) and harvested at day 7 during the differentiation period. The mRNA expression of PPARγ (C), C/EBPα (D) and adiponectin (E) was analyzed by real-time RT-PCR. Values are mean ± S. E. M. of data from three separate experiments (n = 3); each experiment was performed in triplicate. The data were analyzed using analysis of variance (ANOVA) and Duncan multiple range test (P < 0.05). Means sharing a common alphabetical symbol do not differ significantly. (F) Preadipocytes were induced to differentiate with or without FTY-p 0.1 and 1 µM for 2 days. At day 2, the protein levels of p-Akt were analyzed by Western blot. The experiments were repeatedly performed to confirm the results.