Figure 4

Activation of FOXO3a induces Par-4 mRNA expression and apoptosis. (a) PC-3 cells were transiently transfected with FOXO3a (TM) plasmids and an empty vector. After 36 h, cell lysates were prepared, and expression patterns of HA-FOXO3a, pFOXO3a (Ser253), Par-4, BAX, Bcl-2, and p27 were analyzed by western blot analysis. (b) RT-PCR showing the fold increment in Par-4 levels with TM-FOXO3a plasmid overexpression. (c) Effect of FOXO3a-TM on Par-4 promoter activity. (d) Lysates showing the activation of pro-apoptotic proteins Cl-PARP, Caspase-9, and GAPDH/actin was analyzed by western blotting. (e) PC-3 cells were transfected with TM-FOXO3a expression plasmid and stained with annexin-FITC and PI nuclear stain and scored for apoptosis analysis. Significant difference from control values was indicated at P<0.05 (Student's T-test). *P<0.05 and **P<0.005