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. 2015 Jul 1;290(33):20438–20447. doi: 10.1074/jbc.M115.657379

FIGURE 4.

FIGURE 4.

DHA stimulates GLUT4 translocation in AMPK dependently. A, myoblasts stably expressing myc-GLUT4 L6 were differentiated for 7 days. They were pretreated with DHA for 1 h and then treated with insulin for 15 min. Cell surface expression of GLUT4-myc was detected using antibody-coupled colorimetric absorbance assay. *, p < 0.05 compared with control. B, differentiated cells were pretreated with 2 μm compound C and then with DHA for 1 h. Cell surface expression of GLUT4-myc was detected using antibody-coupled colorimetric absorbance assay. *, p < 0.05 compared with control. C, differentiated cells were transfected with AMPKα2 for 2 days and then with DHA for 1 h. Cell surface expression of GLUT4-myc was detected using antibody-coupled colorimetric absorbance assay. *, p < 0.05 compared with control. D, differentiated cells were pretreated with 2 μm STO-609 and then with DHA for 1 h. Cell surface expression of GLUT4-myc was detected using antibody-coupled colorimetric absorbance assay. *, p < 0.05 compared with control cells. E, differentiated cells were transfected with CaMKKβ for 2 days and then with DHA for 1 h. Cell surface expression of GLUT4-myc was detected using antibody-coupled colorimetric absorbance assay. *, p < 0.05 compared with control. F, total mRNA was extracted from DHA-treated C2C12 cells, and RT-PCR was performed using GLUT4-specific primers. PCR products were visualized under ultraviolet light. The results are representative of four independent experiments. *, p < 0.05 versus basal conditions.