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Figure 3


Fig. 3. Rabip4-positive compartments are accessible to internalized Glut 4. 3T3-L1 adipocytes expressing GFP-Rabip4 and DsRed-Glut 4-myc were treated with insulin (100 nM, 20 minutes) and incubated with anti-myc mAb for 2 hours at 4°C. Cells were then extensively washed and either fixed (A) or incubated for 1 hour at the following temperatures: 15°C (B), 20°C (C) or 37°C (D). After fixation and permeabilization, anti-myc mAb were revealed using Cy5-coupled anti-mouse antibodies. Images corresponding to GFP-Rabip4 (in green), internalized anti myc mAb (in blue), the merge of these two images and DsRed-Glut 4-myc (in red) are shown. Colocalization of GFP-Rabip4 with internalized anti-myc mAb (internalized Glut 4) results in a light blue color. Asterisks indicate GFP-Rabip4-positive structures that also contain anti-myc mAb. Bar, 1 µm.





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