Figure 1
From: Regulation of the Intranuclear Distribution of the Cockayne Syndrome Proteins
Regions of CSB predicted to contribute to intracellular localization. (A) Schematic of the domain structure of CSB WT (full-length CSB) and the CSB fragments used in this study. Eight CSB fragments, all of which include the ATPase domain (residues 507-1009, red), are shown. Regions of interest in CSB are: grey: NLS1: 467-481, NLS2:1038-1055, NLS3: 285-354; and orange: NoLS1: 302-341, NoLS2: 1189-1243, NoLS3: 1321-1343. (B) Nucleolar localization of CSB WT in HeLa cells. Cells were prepared following transfection of N-terminally GFP-tagged CSB WT and then visualized for GFP and stained with B23 (nucleophosmin) as a nucleolar localization marker or DAPI as a nuclear marker. Shown are representative images of GFP, B23 or DAPI in HeLa cells. Bar; 10 µm. (C) Localization pattern of CSB fragments in HeLa cells. Shown are representative images of GFP alone, GFP-tagged CSB WT or the eight N-terminally GFP-tagged CSB fragments in live cells. Bar; 10 µm. (D) Quantification of subcellular distribution profile of CSB fragments. The percentage of distribution of CSB fragments into nucleolus (green), nucleoplasm (red) or cytoplasm (blue) was determined from several live cell images (n = 37–70) from (C). Error bars indicate standard deviation (SD).
