EMT status of breast cancer cell lines and CTCs. Cytospin preparations of cells stained for vimentin and K are placed along an axis with increasing vimentin/K (vim/K) ratios and EMT status. In the upper part of the figure are shown representative images of MCF-7, MDA.MB231 and Hs578T cells double stained for vimentin and K and a merge image of EGF treated MCF-7 cells stained for K (green), vimentin (red) and Topro (blue). Numbers shown below the images indicate the mean values of vim/K ratios calculated for each cell line. Numbers next to individual MCF-7 cells undergoing EMT are vim/K values measured for the indicated cells. Characteristic images from CTCs with different vim/K ratios and CD45 staining are presented in the lower part of the figure. Note the absence of CD45 staining in all CTCs and the presence of CD45 positive PBMCs (asterisks). Numbers shown inside the images, indicate the relative fluorescence intensity measured for each demonstrated marker, whereas numbers outside the images represent the vim/K ratios of the CTCs show.