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Fig. 8. Expression of a mutated form of Rabip4 unable to bind Rab4 affects Glut 4 trafficking between early endosomes and its storage compartment. 3T3-L1 Rabip4 
507-517 Tet-off adipocytes were cultivated for 2 days in the presence (a-c), or absence of tetracycline (d-f) to induce the expression of Rabip4 507-517. Cells were serum deprived before a 20 minute treatment with 100 nM insulin. After extensive washes in DMEM containing 0.5% BSA, the adipocytes were incubated for 1 hour at 37°C. They were then fixed, permeabilized and used for immunofluorescence analysis. Glut 4 was detected using anti-Glut 4 goat polyclonal antibodies followed by FITC-coupled anti-goat antibodies (a,d), whereas TfR were detected with anti-TfR mAb followed by Texas-Red-coupled anti-mouse antibodies (b,e). The merged images are shown in panels c and f. The dotted square inserts show cytoplasmic vesicles with Glut 4 labeling in green and VAMP2 labeling in red. Bar, 1 µm.
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