Figure 5. Aurora A regulates FoxO1 expression at the transcriptional level in a p53-dependent manner. (A) Hep3B cells were co-transfected with a combination of indicated plasmids, incubated for 48 h, and western blot analysis was performed. (B) FoxO1 mRNA half-life was determined by actinomycin D treatment. Vector control and Aurora A shRNA cells were treated with actinomycin D (10 μg/ml) for the indicated time points. FoxO1 mRNA levels were measured by real-time qRT-PCR. (C) For luciferase reporter assay, vector control and Aurora A-knockdown cells were transfected with luciferase reporter plasmid containing FoxO1 promoter. pGL3 vector was used as a negative control. Data shown are mean ± SD of three independent experiments. *p < 0.01. (D) HepG2 cells were transfected with a combination of the Aurora A shRNA and RNAi-resistant Aurora A variant in the presence of luciferase reporter plasmid containing FoxO1 promoter. pGL3 vector was used as a negative control. Data shown are mean ± SD of three independent experiments. *p < 0.05. (E) Hep3B cells were co-transfected with a combination of the indicated plasmids in the presence of luciferase reporter plasmid containing FoxO1 promoter. pGL3 vector was used as a negative control. Data shown are mean ± SD of three independent experiments. *p < 0.05.