Time-lapse microscopy of cells expressing vac17-S222A-GFP were labeled with FM4-64 (vacuole) and imaged every 60 s. In cells expressing the vac17-S222A-GFP mutant that could not be phosphorylated, the vacuole and vac17-S222A-GFP traveled from the bud tip to the mother-bud neck. Representative time lapse from n = 3. Image frames were acquired every 60 s; video is displayed at three frames per second.
