Figure 5. ERK2-NES was localized in the cytoplasm when activated by MEK1. For fluorescence studies, hepatocytes were transfected with ERK2-NES-GFP or ERK2-AAA-GFP combined with MEK1wt together with Racwt. A) Confocal fluorescence images of control and 5 min of EGF-stimulated cells with ERK2-NES-GFP (a, b), and control and 5 min of EGF-stimulated cells expressing ERK2-AAA-GFP (c, d). B) Mean ERK2wt-GFP cytoplasmic localization in control and EGF-stimulated cells. Data presented are the mean ± SE of 3 independent experiments. C) Confocal fluorescence images of control and 5 min of EGF-stimulated cells with ERK2-NES-GFP in the presence of LeptomycinB (a, b). D) Western immunoblotting on hepatocytes transfected with ERK2wt-GFP or ERK2-NES-GFP combined with either MEK1wt or MEK2wt with and without EGF with an antibody recognizing T-E-Y-phosphorylated ERK.