Porcine hepatocyte isolation and reversible immortalization mediated by retroviral transfer and site-specific recombination

Figure 5 Scheme of reversible immortalization. The primary porcine hepatocytes were immortalized by transfer of an oncogene (SV40T). After expansion of the immortalized cells, Cre/loxP recombination was performed to remove the oncogene (SV40T) and the cells reverted to their pre-immortalized state. (A-C) Double immunofluorescence of immortalized hepatocytes stained with DAPI (blue), which binds, together with monoclonal antibody anti-SV40T revealed with texas red-antibody conjugate (red). Blue and red fluorescence merged as purple. SV40T expression is revealed by intense staining of the cell nucleus.