Real time analysis of GFP–Psc1 motility in COS-1 cells

Copyright information:

Taken from "A family of RS domain proteins with novel subcellular localization and trafficking"

Nucleic Acids Research 2005;33(4):1309-1322.

Published online 1 Mar 2005

PMCID:PMC552957.

© The Author 2005. Published by Oxford University Press. All rights reserved

GFP–Psc1 transfected COS-1 cells were analysed 10 h post transfection by confocal microscopy. Images were captured at 15–30s intervals. Cells were stained with Hoechst 33342 to identify the nucleus and maintained in fresh growth medium at 37°C for the timecourse of the experiment. () Motility of Psc1 nuclear speckles. The arrow indicates speckle motility and budding across the nucleus. The lower arrow in panel 4 indicates a speckle originated from a budding event. Panels 1–6 were captured at 0, 30, 90, 270, 420 and 540s, respectively. (B–D) Motility of Psc1 cytospeckles. () Random and stationary cytospeckle motility. The cytospeckle indicated by the arrow in panels 1–12 shows a change of direction in panel 10. Upper arrow in panel 12 shows a stationary cytospeckle. Panels 1–12 were captured at 0, 15, 30, 45, 75, 90, 105, 120, 150, 165, 210 and 225, respectively. () Directed motility and fusion of cytospeckles. The cytospeckle indicated by the arrow moves directionally away from the nucleus and fuses with distant cytoplasmic speckles. Panels 1–5 were captured at 0, 630, 780, 900 and 930s, respectively. () Motility of Psc1 cytospeckles, (arrow, panels 1–8). Panel 4 shows the speckle move slightly away from the nucleus before apparent nuclear translocation in panels 5–8. Panels 1–8 were captured at 0, 15, 45, 90, 105, 120, 135 and 150s, respectively. GFP was visualized by direct fluorescence under excitation at 480 nm using confocal microscopy. Size bars represent 5 μm.