Figure 2

Without the context of the full length protein, NES2 can redirect GFP to the cytoplasm, while NES1 function depends on its positioning vis-à-vis GFP. (A): Sequence alignment of a consensus NES sequence with Tax1 NES and Tax2 putative NES. (B): HeLa cells were transiently transfected with NES-EGFP and GFP-NES plasmids. Twenty-four hours after transfection, the cells were washed with PBS, fixed with 4% paraformaldehyde, mounted with DAPI-containing mounting medium and visualized with a Zeiss Axioplan 2 imaging microscope X40 using a Zeiss Axiocam HRc (color) camera and the Zeiss Apotome software. Images of cells that are representative of the entire population are shown. (C): Western-blot analysis of cytoplasmic and nuclear cell fractions. 293T cell fractions were subjected to electrophoresis on a 10 % TG gel and probed with a GFP antibody. The western-blot results are representative of four independent experiments.