Time-lapse imaging of dividing BRCC36-silenced cells, which were rescued by DUB-inactive mutant CFP-BRCC36res QSQ, based on Fig. 5. H2B-GFP/α-tubulin–mRFP HeLa cells were cotransfected with BRCC36-siRNA and CFP-BRCC36res QSQ plasmids. Images were analyzed by time-lapse confocal microscopy using a laser-scanning confocal microscope (40× oil objective, Plan-Apochromat, NA 1.4; Eclipse Ti-E; Nikon) 36 h after transfection. Frames were taken every 5 min for 24 h at 37°C.
