Fig. 3. NRAGE affects Che-1 cellular localization. For co-expression experiments, NIH3T3 cells were doubly transfected with EGFP-NRAGE and Myc-Che-1 or one of the two Myc-Che-1 deletion mutants (Myc-Che-1-C, Myc-Che-1-C). After 48 hours, cells were fixed and immuno-stained with antibody against Myc-epitopes as described in Materials and Methods. Sub-cellular distribution and colocalization of Myc-tagged proteins (red) with EGFP-NRAGE (green) were analysed using fluorescence microscopy. No overlapping staining was observed in cells co-expressing Myc-Che-1-C and EGFP-NRAGE fusion proteins. Note that, EGFP-NRAGE expression affects the nuclear localization of Myc-Che-1.