Live-cell imaging of GLUT1 and GLUT4 after simultaneous release from the ER: example 1. HeLa cells expressingthe ER Ii-hook fused to streptavidin were transfected with HA-GLUT1-SBP-mCherry (GLUT1, red) and HA-GLUT4-SBP-GFP (GLUT4,green). The intracellular traffic of GLUT1-mCherry and GLUT4-GFP was simultaneously tracked in a single cell for 1 h after biotinaddition concurrently released them from the ER. Upon ER exit, both GLUT1 and GLUT4 accumulate in the perinuclear region ofthe cell (yellow), and then highly mobile GLUT1 vesicles (red) become visible toward the cell periphery while GLUT4 remainsperinuclear. The time-lapse video was acquired on a spinning-disk microscope at 2 frames/s. Stills from this video are shown inFig. 2 A. Scale bar: 10 μm.
