To do so, PCa cells (LNCaP and DU145) were transfected with plasmids encoding for either empty vector, Flag-tagged FOXO1 WT, or a phospho mutant (S75A or S75D), and analyzed the subcellular localization of the proteins by immunofluorescence using an anti-Flag antibody. Here, FOXO1 is linked to posterior cortical atrophy.