Cdk5 promotes an AKT-independent nuclear export of FOXO1. A, B, Primary cortical neurons were transfected with Flag-FOXO1 together with GFP, GFP-p25/Cdk5, GFP-p35/Cdk5, GFP-p25/Cdk5-KD, or GFP-p35/Cdk5-KD on DIV 3. On DIV 7, the neurons were treated with vehicle control (DMSO) (A) or LY294002 (B) for 16 h. After treatments, the cells were fixed to measure the subcellular localization of FOXO1 by Flag-immunofluorescent staining. Representative images are shown on the left: green represents GFP; red represents Flag; blue represents DAPI. Quantification of subcellular localization of FOXO1 is shown on the right. C, D, Primary cortical neurons were transfected with Flag-FOXO1-AAA or Flag-FOXO1-AAA-S249A together with GFP, GFP-p25/Cdk5, GFP-p35/Cdk5, GFP-p25/Cdk5-KD, or GFP-p35/Cdk5-KD. On DIV 7, the cells were fixed to measure the subcellular localization of FOXO1 by Flag-immunofluorescent staining. Representative images are shown on the left: green represents GFP; red represents Flag; blue represents DAPI. Quantification of subcellular localization of FOXO1 is shown on the right.