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Fig. 6. Time course of fluorescent transferrin uptake and recycling and its association with SCAMP-containing compartments. (A-P) Alexa 594-transferrin (100 µg/ml) was bound to NRK cells at 0°C and uptake was initiated by warming to 37°C for the specified time. Whole cell images of uptake after 1 minute (A) and 3 minutes (B). (C,D) Deconvolved images 10 seconds (C) or 20 seconds (D) after uptake in cells stained for clathrin. (E-P) Deconvolved images comparing internalized transferrin at indicated times to the staining of SCAMP1-4. (E-I) portions of peripheral areas are shown. (M-P) portions of perinuclear areas are shown. (Q-T) To follow recycling carriers, cells were labeled with Alexa 594-transferrin for 10 minutes and chased for 60 minutes at 37°C. Deconvolved sections of perinuclear areas comparing transferrin and the staining of the SCAMPs are shown. Bar, 10 µm (B); 2 µm (D,T).
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