Movie 6

example of streptavidin recovery in nanotube upon fusion with a GUV containing CHMP2B-ΔC.
The movie shows four sequential bleaching and recovery events on a nanotube with streptavidin bound to the inner leaflet. Streptavidin, lipids and CHMP2B- ΔC are depicted in yellow, magenta and green, respectively. In the first three FRAP events, no CHMP2B cluster is visible at the neck, and streptavidin recovers very quickly. At the end of the third recovery, a CHMP2B cluster happens to localize at the neck region. After a subsequent bleaching, no recovery of streptavidin is observed. However, as soon as the CHMP2B cluster moves away from the neck, quick recovery occurs. Frame rate: 5fps. The movie has been paused before and after each bleaching event to better visualize the fluorescence of the nanotube. The plot in the lower part of the movie displays the quantification of streptavidin fluorescence in the nanotube during the recovery, normalized for the fluorescence before each bleaching event. Green and orange dots indicate frames in which the CHMP2B cluster was localized at the neck or not, respectively.