Figure 4

The presence of leucine 188 restores NES1 function within the context of a NES1-EGFP protein. (A): Western-blot analysis of the different EGFP and NES-EGFP proteins. 293T cell lysates (70 μg) were subjected to electrophoresis on a 10 % TG gel and probed with GFP or β-tubulin antibodies. (B): HeLa cells were transiently transfected with the different NES-EGFP plasmids. Eighteen hours post transfection, transfected cells were washed, fixed, mounted with DAPI-containing 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): 293T nuclear and cytoplasmic 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.