Role of importin α/β nuclear import pathway in the nuclear distribution of C-Tpr. Digitonin-permeabilized NRK cells were incubated with 8 pmoles Cy5-C-Tpr (Panels A, C, E, G) or 8 pmoles Cy3-BSA-NLS (coupled with a synthetic peptide comprising the NLS of SV40 T-antigen; Panels B, D, F, H) and nuclear import was conducted with recombinant factors as described in Materials and Methods. Panels A, B: nuclear import of Cy5-C-Tpr and Cy5-BSA-NLS performed at 30°C; panels C, D: importin α was omitted; panels E, F: importin β was omitted; panels G, H: Cy5-C-Tpr and Cy3-BSA-NLS competed with 10-fold molar excess of unlabeled cargo. Samples were visualized by confocal fluorescent microscopy.
Ben-Efraim et al. BMC Cell Biology 2009 10:74 doi:10.1186/1471-2121-10-74