FIG. 10.

Nuclear recruitment of FOXO3a is associated with cytoplasmic accumulation of active CYB5R3. MRC-5 cells were transfected with expression plasmids encoding human FOXO3a mutant fused with the ligand-binding domain of estrogen receptor (p-FOXO3a-A3-ER). Control transfection was carried out with empty plasmid. (A) Nuclear localization of activated FOXO3a was determined by western blot and immunofluorescence after a 24-h stimulation with 4-OHT (100 nM). (B) Enzymatic activity of CYB5R3 and immunoblot analysis of CYB5R3 and FOXO3a was performed in cytoplasmic fractions. Membranes were reprobed for α-TUBULIN, which was used as a loading control. Means±SD of at least three different experiments are represented. **p<0.001. 4-OHT, 4-hydroxytamoxifen.