Fig. 4.

The mTOR inhibitor, rapamycin, suppresses adipogenesis of C3H10T1/2 cells.

A) Confluent CH3H10T1/2 cells were induced to differentiate by addition of 10% foetal calf serum (FCS) supplemented with adipogenic cocktail (IID), in the presence or absence of the mTOR inhibitor, 10 μM rapamycin. After 5 days cells were fixed with formalin and stained with Oil Red O to detect neutral lipid accumulation. Representative micrographs from an experiment carried out on three separate occasions with similar results are shown.

B) Confluent C3H10T1/2 cells were transfected with PPARγ luciferase gene reporter construct, together with control Renilla luciferase vector and then stimulated for two days with 500 μM metformin or 10 μM rapamycin, in the presence or absence of IID. Cell extracts were then prepared and luciferase activity was measured using a dual luciferase reporter assay. Luciferase activities from three separate experiments are shown as means ± SEM. Significant increases in PPARγ activity are indicated ***, p < 0.001, as are significant decreases in PPARγ activity, #, p < 0.05, relative to IID-stimulated cells (n = 3).