The cartoon shows the research strategy used to experimentally validate the predicted subcellular localization of proteins. Genes of interest were cloned into pEGP-N1 vector as GFP fusions. These GFP-tagged genes along with corresponding location-specific RFP-tagged gene markers, were transiently co-transfected using liposome-mediated method into two normal breast and breast cancer cell lines. Following 24 hours of transfection and protein expression, subcellular localization was determined using live cell imaging/confocal microscopy. Protein localizations to each compartment were confirmed by observing the colocalization of GFP- and RFP-tagged proteins that gives yellow color up on merging green and red images.