Fig. 2

Sub-cellular localization of the MDM2-splice variants. (a) MCF-7 cells transfected with pCMV-GFP (GFP-control), MDM2-FL, -A, -B and -C evaluated by indirect immunofluorescence for determination of cellular localization of the proteins. Top lane shows the nucleus by Hoechst staining (blue), second lane shows the cells expressing GFP (green), third lane shows the AlexaFlour 647 MDM2 antibody (Life Technologies, red), and lastly, an overlay of the tree previous pictures. (b) Percentage of cells with exclusively nuclear (dark blue bars), nuclear and cytoplasmic (blue bars) or exclusively cytoplasmic (light blue bars) localization of the MDM2-A, -B and -C. For each of the differently transfected cell samples the sub cellular localization was determined for 50 transfected cells with each of the constructs. The experiment was repeated in triplicate, with three independent transfections. Cells were counted by three independent investigators blinded to sample identity and each other’s results