Fig. 7. Analysis of domains in SF3a66 required for nuclear and subnuclear localization. (A) Schematic representation of recombinant SF3a66 proteins fused to GFP. Amino acids present in, or deleted from the recombinant proteins are given in parentheses after the names of the proteins. Conserved regions are indicated above the diagram and numbered according to the amino acids in full-length SF3a66. (B) GFP-tagged 3a66-N, -C and -Zn were transiently expressed in HeLa cells and visualized by fluorescence in living cells. (C) HeLa cells were transiently transfected with plasmids encoding GFP-SF3a66 proteins as indicated. Fluorescence was monitored after fixation of cells with cold methanol. Cells in (d) were immunostained with anti-p80-coilin (middle panel); the right panel represents the computer-generated overlay. (D) HeLa cells were transiently transfected with GFP-3a66-Zn. Proteins (as indicated) were visualised after fixation of cells with cold methanol. SF3a60 was visualised by immunofluorescence with anti-3a60 and AMCA-coupled anti-rabbit antibodies, p80-coilin with a mouse anti-p80-coilin and Texas Red-coupled anti-mouse antibodies. Arrows point to CBs. Bar, 10 µm.