Figure 2
Mapping of the nuclear localisation signal of EWS-Oct-4B to the POUB DNA-binding domain. (A) Subcellular localisation of EWS-Oct-4B. Cells (293T) grown on coverslips were transfected with mammalian expression vectors encoding (a) EGFP or (b) EWS-Oct-4B-EGFP. The subcellular localisation of the EWS-Oct-4B protein was determined by monitoring the location of the green fluorescence. (B) Schematic diagram showing EWS-Oct-4B truncation mutants of GST-EGFP fusion proteins. Subcellular localisation of the indicated truncation mutants was determined by monitoring the location of green fluorescence, and is indicated as N (nuclear localisation) or C (cytoplasmic localisation). (C) Subcellular distribution of EWS-Oct-4B deletion mutants. 293T cells were grown on coverslips under low-density conditions and transfected with expression plasmids for the indicated GST-EGFP-EWS-Oct-4B deletion mutants. The cells were fixed with an acetone/methanol mixture and EGFP was analysed by fluorescence microscopy.
