Fluorescent images for the dynamic behaviors of inner nuclear membrane proteins and NLS-BSA during in vitro nuclear envelope reconstitution reaction

Fluorescent images of LBR-YFP, CFP-emerin and Cy3-NLS-BSA during the nuclear envelope reconstitution reaction in mitotic semi-intact cells. 

Semi-intact cells prepared from HeLa cells stably co-expressing LBR-Venus and SECFP-emerin by digitonin treatment. Images were captured just after the addition of a reaction mixture to digitonin-permeabilized semi-intact cells and at 5-min intervals throughout the reactions for 60 min (t01~t013) at 30 °C with a DeltaVision microscope. The reaction mixture was composed of an ATP regeneration system, GTP, mitotic cytosol, cyclin-dependent kinase 1 inhibitor (RO-3306) and Cy3 labeled bovine serum albumin tagged with SV40 T-antigen nuclear localization signal (Cy3-NLS-BSA) as a transport indicator. 

The file names indicate fluorescent labeled proteins (LBR-Venus , LBR-YFP; SECFP-emerin, CFP-emerin; Cy3-NLS-BSA), mitotic phases (metaphase, anaphase, telophase) of semi-intact cells, and the incubation time point (t01~t13) of each image. The merged images (merge) show images of SECFP-emerin (blue), LBR-Venus (green) and Cy3-NLS-BSA (red).