FRAP of G3BP1-GFP and GFP-ATXN2L within G3BP1-induced lysate granules. Video corresponds to stills and FRAP curve shown in Fig. 2, H and I. Lysates from stable G3BP1-GFP or KI GFP-ATXN2L U2OS cells were induced by increasing the concentration of G3BP1 by 20 µM using purified G3BP1. Imaging was performed using time-lapse spinning-disk confocal microscopy of granules settled at the surface of the imaging chamber. Arrows indicate lysate granules on which FRAP was performed. Images were captured every 0.2 s for 35 s total. Photobleaching with 488-nm laser occurred 2 s into capture. Frames per second = 15.
