Figure 3. Foxo3 directly targets miR-422a. (A) Subcellular distribution of Foxo3 in SKOV3 cells, as investigated by fluorescence in situ hybridization (FISH) assay. Scale bar = 25 μm. (B) Subcellular distribution of Foxo3 in SKOV3 cells, as investigated by subcellular fractionation assay. GAPDH and U6 were used as reference genes for the nuclear and cytoplasmic, respectively. The data were expressed as cytoplasmic/nuclear ratio. (C) The putative binding sequence of miR-422a of 3’UTR of Foxo3. The prediction was obtained from StarBase 2.0. (D) Luciferase reporter assay in HEK293T cells co-transfected with miR-422a mimics or mimics a negative control plus reporter vector containing wild-type or mutant miR-422a binding sequence 3′-UTR of Foxo3. (E) The expression of miR-422a in SKOV3 and control cells. (F) The expression of miR-422a in SKOV3 cells with Foxo3 knockdown or overexpressing. Data were expressed as means ± standard deviation. For each treatment, three replicates were used. **p < 0.01, ***p < 0.001.