Mapping the putative nuclear localization signals (NLS) of YB-1. A. Diagram of DsRed-tagged abbreviated residues corresponding to putative NLS. The diagram summarizes the four putative NLS and their relative localization within the full-length YB-1 protein. B. Subcellular localization of putative NLS in RMC. Cells were transfected with expression vectors encoding for DsRed-tagged potential NLS of YB-1 and CFP, respectively. The subcellular localization was determined using confocal laser scanning microscopy. The cellular morphology was visualized by co-transfection with CFP, that enriches within the nucleus. The histograms provide quantification of the staining pattern. 100 transfected cells were assessed for each construct. C. Mutational analyses of the NLS-2 (PNLS185) motif. Diagram on DsRed-tagged mutated PNLS185 expression vectors generated to express the wild-type or mutated sequences (N: nuclear localization). D. Localization of the mutated PNLS185 expression proteins in RMC. Cells were transfected with DsRed-tagged expression vectors encoding for mutated PNLS185 and CFP, respectively. The histograms provide quantification of the staining pattern. 100 transfected cells were assessed for each construct.