Movie 7.

Representative confocal microscopy time lapses of fluorescence recovery after photobleaching (FRAP) experiments demonstrating differential recovery of EmRab7 in pH neutralization conditions, and RILP/V₁G₁ binding mutants. The following stable NRK cells were treated with the following conditions and subjected to photobleaching: EmRab7^WT-DMSO (top left), EmRab7^WT-LLOMe (top right), EmRab7^WT-NH₄Cl (middle left), EmRab7^WTBafA1 (middle right), EmRab7^L8A-DMSO (bottom left), EmRab7^L8A-DMSO (bottom right). A 33.0625μm² area was bleached for 10 seconds using a 488-laser line. Images were captured for 1 minute prior to bleach, and 4 minutes post-bleach at a rate of 10 seconds per frame. The bleached and unbleached control area used for normalization are indicated with yellow boxes. DMSO, LLOMe (1mM), and NH4Cl (20mM) treatments were for 10 minutes prior to imaging. BafA1 (200nM) treatment was for 1 hour. Scale bar = 20μm, frame rate = 4fps.