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. 2016 Jun 27;6:28648. doi: 10.1038/srep28648

Figure 3. Dexras1 is required for IGF-1-induced ERK activation, but not for AKT activation, in adipogenesis.

Figure 3

(a) pcDNA3-Dexras1-FLAG overexpressing 3T3-L1 cells were incubated with various concentrations of insulin or IGF-1 for 30 min. Immunoblots are a representative image of three independent experiments, and the activation levels of ERK or AKT were measured and plotted. Data are presented as the mean ± s.d. **denotes P < 0.01. (b) 3T3-L1 cells transfected with pcDNA3-Dexras1-FLAG were differentiated with various concentrations of insulin or IGF-1 in the presence of IBMX. Oil-red-O staining was performed on day 8. (c) Cells were transfected with pcDNA3-Dexras1-FLAG or pcDNA3-∆223-276-FLAG. After 30 min of pretreatment with the indicated reagents (U, U0126; PD, PD98059; W, wortmannin; or LY, LY294002) cells were incubated with IBMX and insulin (1 μg/ml) for 10 min. ERK and AKT activation was evaluated by immunoblot analysis. (d) Dexras1-overexpressing 3T3-L1 cells were transfected either with scramble or IGF-1R siRNAs, and then stimulated by insulin (1 μg/ml) for 1 h. Cells were fixed and subjected to immunofluorescence analysis with antibody against FLAG (green). Nuclei were stained with DAPI and fluorescence was visualized by confocal microscopy. Scale bar = 50 μm.