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. 2013 Apr 16;62(5):1519–1526. doi: 10.2337/db12-1066

FIG. 7.

FIG. 7.

ATP-dependent GLUT4myc translocation requires VAMP2 and reduces GLUT4myc endocytosis. A and B: Primary myotubes were transiently cotransfected with GLUT4myc-eGFP and TeTx cDNA for 24 h and then stimulated with 100 μmol/L ATP for 10 min. C: GLUT4myc endocytosis in L6-GLUT4myc cells. Surface GLUT4myc was labeled with anti-myc antibody, followed by extensive washing with PBS. The surface-labeled GLUT4myc was then allowed to internalize by rewarming for different times in presence or absence of the respective stimulus. At the indicated times, the amount of GLUT4myc remaining at the cell surface was determined and expressed as a percentage of the cell surface level at 0 min of endocytosis. D: GLUT4myc re-exocytosis in L6-GLUT4myc cells. Cells were first stimulated with 100 nmol/L insulin for 10 min to induce GLUT4myc translocation. Surface GLUT4myc was labeled with anti-myc antibody, followed by extensive washing with PBS, and then warmed to 37°C for 2 h to induce endocytosis in α-minimum essential medium. L6-GLUT4myc cells were then stimulated with exogenous ATP or insulin for 10 min. Subsequently, cells were again cooled to 4°C, and antibody-laden GLUT4myc was determined at the cell surface. Values are the mean ± SD. **P < 0.001, *P < 0.01 vs. basal group. †P < 0.01 with respect to unstimulated control. ††P < 0.001 with respect to ATP-stimulated cells. Data are from at least four independent experiments.