Tyrosine Phosphorylation in the C-Terminal Nuclear Localization and Retention Signal (C-NLS) of the EWS Protein
Figure 2
(a) Schematic representation of the N-terminally tagged GFP-Zf fusion protein. 6His-GFP is fused to the EWS mutant protein (aa 525–656). (b) Representative examples of the subcellular localization (all cells expressing the construct are showing the indicated localization) of GFP-Zf, GFP-Zf(Y656A), and GFP-Zf(Y656A)-SV40NLS (in green). Nuclei are shown by DAPI staining (in blue). Bars, 15 μm. (c) Phosphorylation analysis of GFP-Zf and GFP-Zf(Y656A) using antiphosphotyrosine antibodies. GFP-Zf and GFP-Zf(Y656A) fusion protein bands are detectable at ~40 kDa. The additional phosphorylated protein band detected at ~55 kDa was identified as p54nrb.
