Figure 5. FOXO4-DRI causes nuclear exclusion of active p53 and induces apoptosis in senescent TM3 Leydig cells. (A) Immunofluorescent staining showing increased levels of Ser15-phospho-p53 in the nucleus of senescent TM3 Leydig cells. Note that Ser15-phospho-p53 foci are excluded from the nucleus after treatment with 25 mM FOXO4-DRI for 3 days. Scale bar: 25 μm. (B) CCK8 assays showing that FOXO4-DRI decreases the viability of senescent TM3 Leydig cells. (C, D) Annexin V-FITC/PI apoptosis assays showing that FOXO4-DRI increases the apoptosis rate among senescent TM3 Leydig cells. Data presented are representative of three independent experiments. Data depict the mean ± SD. *P<0.05. NS, nonsignificant.