Fig. 2

EN2 expression via immunofluorescent staining of platinum-sensitive/−resistant paired serous ovarian cancer cell lines and controls. (a) EN2 fluorescent staining (green) was present in the cytoplasm of the serous cell lines, particularly evident in those that had been permeabilised. There was no EN2 staining evident in the permeabilised negative control of each cell line, where only secondary antibody was added (inset). There was no EN2 staining evident in the fibroblast cell line, whilst there was strong cytoplasmic staining in the melanoma A375M positive control cell line. TOPRO staining (blue) identified the cell nucleus and WGA staining (red) identified the cell membrane. Co-localisation of cell membrane WGA and EN2 protein staining (yellow) can be seen in some of the non-permeabilised cells (blue arrows). (b) Intensity profiles for each label along the length of the white arrow are shown in a PEA2 cell, and a PEO4 cell (c), demonstrating the absence of EN2 in the nucleus, but close association with the cell membrane. The coloured peaks represent the cell membrane, nucleus, presence of EN2, and co-localisation of EN2 with the cell membrane, as detailed above