FRAP of transfected GFP-NPM1 in the nucleolus and NPM1-induced lysate granules. Video corresponds to stills and FRAP curves shown in Fig. 4, B and C. U2OS cells were transiently transfected with GFP-NPM1. Intact transfected cells were imaged to monitor mobility of GFP-NPM1 in the nucleolus. Lysates prepared from transfected cells were induced by increasing the concentration of NPM1 by 15 µM using purified NPM1. Imaging was performed using time-lapse spinning-disk confocal microscopy of granules settled at the surface or cells adhering to the surface of the imaging chamber. Arrows indicate nucleolus and NPM1-induced lysate granule on which FRAP was performed. Images were captured every 0.2 s for 60 s total. Photobleaching with 488-nm laser occurred 2 s into capture. Frames per second = 30.
