Figure 7

The tyrosine-based motif in position 23 allows the chimera to follow a retrograde route from endosomes to TGN. HeLa cells were transfected with A wild type, B L3S mutated or C Y23S mutated chimeras. Forty-eight hours after transfection, cells were treated with cycloheximide for 2 h. Chimeras present on cell surface were stained with the anti-CD25 antibody at 4°C for 1 hour and cells were then shifted at 37°C for another hour. After fixation, internalized anti-CD25 was revealed using FITC-conjugated secondary antibodies, and MPR46 was revealed as in figure 1.