Movie S2

Mroh1-GFP is not recruited to new endosomes
Cells co-expressing PH-CRAC-mCherry and Mroh1-GFP were imaged by spinning disk confocal microscopy at a frame interval of 2 sec. Large macropinosomes were formed, showing the presence of PH-CRAC. These were internalized into the cell to form early endosomes. In the example shown, one early endosome that still has its PH-CRAC coat can be seen to jostle for several minutes in the vicinity of several different vesicles that are labeled with Mroh1-GFP, but no Mroh1-GFP signal appears on the early endosome.