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Figure 6


Fig. 6. Characterization of a CRM1 specific STAT2 NES element. (A) Sequence of human STAT2 between aa position 690 and 759. Positions of STAT2 C-terminal deletion mutants are marked by arrows and aa mutated to alanine are underlined. (B) Subcellular localization of STAT2-{Delta}754-EYFP (a), STAT2-{Delta}703-EYFP (b), STAT2-L733A-EGFP (c), STAT2-L747A-EGFP (d) and STAT2-LL737/741AA-EYFP (e) was monitored in untreated C243 cells by confocal analysis after transient transfection. Nucleocytoplasmic distribution (f) of STAT2-LL737/741AA-EYFP was compared in C243 cells (left) before and (right) after LMB treatment for 2 hours. The nuclear region is enclosed by the blue and red lines inside the graph of the staining profile. (g) Localization STAT2-LL737/741AA-EYFP was monitored after its transfection into U2A cells. (C) NIH3T3 cells stably expressing STAT1-CFP were transiently transfected with STAT2-LL737/741AA-EYFP expression plasmid. Subcellular localization of both STATs was determined by fluorescence microscopy in untreated cells and after stimulation with IFN-ß for 90 minutes. IFN was removed and nucleocytoplasmic distribution of STAT2-LL737/741AA-EYFP and STAT1-CFP was monitored 120 min later.





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