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. 2009 Jun 23;94(9):3535–3542. doi: 10.1210/jc.2009-0162

Figure 4.

Figure 4

Inhibition of translocation of GLUT4 and GLUT12 in L6 muscle cells in culture. L6 cells were cultured to near confluence in 35-mm dishes, and then the growth medium was replaced with medium without serum for 1 h. Cells were then incubated at 37 C for another hour in the presence or absence of insulin (10 ng/ml) with concentrations of the PI3-K inhibitor LY294002 at 0, 1, 10, and 100 μm. After harvesting and fractionating the cells, samples from the fractions were subjected to immunoblotting, and digital images were quantified by image analysis software. A, Typical immunoblots; B and C, summary graphs of the quantitative analysis. Portions of both GLUT4 and GLUT12 moved from the LDM fraction to the PM fraction in response to incubation with insulin. At 1 μm LY294002 in the absence of insulin, GLUT4 and GLUT12 were more in the PM fraction than in the absence of LY294002, suggesting a PI3-K agonist effect at the lowest concentration. The translocation induced by insulin was reduced by the presence of LY294002 in a dose-responsive manner over the full range of concentrations tested.