Insulin-stimulated glucose transport is a characteristic property of adipocytes and
muscle cells and involves the regulated delivery of glucose transporter (GLUT4)-
containing vesicles from intracellular stores to the cell surface. Fusion of these
vesicles results in increased numbers of GLUT4 molecules at the cell surface. In an
attempt to overcome some of the limitations associated with both primary and
cultured adipocytes, we expressed an epitope- and GFP-tagged version of GLUT4
(HA–GLUT4–GFP) in HeLa cells. Here we report the characterisation of this system
compared to 3T3-L1 adipocytes. We show that insulin promotes translocation of
HA–GLUT4–GFP to the surface of both cell types with similar kinetics using
orthologous trafficking machinery. While the magnitude of the insulin-stimulated
translocation of GLUT4 is smaller than mouse 3T3-L1 adipocytes, HeLa cells offer a
useful, experimentally tractable, human model system. Here, we exemplify their
utility through a small-scale siRNA screen to identify GOSR1 and YKT6 as potential
novel regulators of GLUT4 trafficking in human cells