Fig. 1.

Insulin-stimulated triacylglycerol (TAG) accumulation in 3T3-L1 adipocytes requires mTORC1 signaling. a 3T3-L1 adipocytes were serum-deprived for 20 h (Control) and then pre-treated without or with rapamycin (20 ng/mL) for 30 min. Cells were stimulated without (Control) or with insulin (INS) (100 nM) in the absence (R) or continued presence of rapamycin (R + INS) for 18 h. Whole cell lysates were analyzed for triacylglycerol content (mg/mg protein) via a colorimetric assay (see “Experimental Procedures”) after normalization for protein content. The graph represents the mean ± SD of triplicate samples from one experiment but is representative of three independent experiments; thus, each bar represents n = 3. Differences between treatment groups were analyzed using ANOVA, and differences were considered statistically different at a level of 0.05. Bars with different letters are statistically significant. b The whole cell lysates analyzed in (a) were resolved on SDS–PAGE and immunoblotted with the indicated antibodies to confirm the expected activation and/or inhibition of mTORC1 signaling