JCS02503 Supplementary Material

Files in this Data Supplement:

• Movie 1 - Movie 1. NRK cell expressing GFP-SCAMP1 labeled 10 minutes with Alexa-594 Tf and chased approximately 90 seconds. Note the extensive colocalization of GFP-SCAMP1 and transferrin and their simultaneous movement. In the lower half near the center, an endosome undergoes upward saltatory movement.

• Movie 2 - Movie 2. NRK cell expressing GFP-SCAMP1 after a 15-minute labeling with Alexa-594 Tf followed by a 40-minute chase. Tf is generally present in endosomes that do not contain GFP-SCAMP1. Toward the left a Tf-containing endosome moves from the bottom of the field to the top and subsequently a GFP-SCAMP1 vesicle is seen to translocate from lower left to upper right.

• Movie 3 - Movie 3. NRK cell expressing GFP-SCAMP1 after a 15-minute labeling with Alexa-594 Tf followed by a 40-minute chase. This movie presents additional examples of separate movement of Tf and GFP-SCAMP1. Endosomes containing Tf but not GFP-SCAMP1 move toward the top of the field. Toward the bottom of the field GFP-SCAMP1-containing vesicles are translocating.

• Movie 4 - Movie 4. NRK cell expressing GFP-SCAMP1 labeled 15 minutes with Alexa-594 Tf and chased 40 minutes. In the center, distinct GFP-SCAMP1 and Tf-filled endosomes appear to be attached and tugging on one another as if trying to separate.