Figure 2. Depletion of Aurora A induces upregulation of FoxO1 expression in HepG2 and SK-HEP-1 cells, and RNAi-resistant Aurora A downregulates FoxO1 expression. (A) Western blot analysis of total FoxO1, phospho-FoxO1 (S256), p21 and Aurora A expression in vector or Aurora A shRNA-expressing HepG2 and SK-HEP-1 cells. (B) Western blot analysis of total FoxO1, phospho-FoxO1 (S256), p21 and Aurora A expression in vector or Aurora A shRNA-expressing Huh-7 and Hep3B cells. Two different Aurora A shRNA were used. Equal loading of protein was confirmed by GAPDH. (C) Nucleotides substituted (base changes are underlined) in the vector expressing the RNAi-resistant Aurora A. (D) HepG2 cells were transduced with pLKO.1 or Aurora A shRNA lentiviral particles. At 48 h after the transduction, cells were transfected with RNAi-resistant Aurora A containing or empty vector. Protein lysates were extracted 48 h later and followed by western blot analysis with the indicated antibodies. (E) Cell proliferation in control and Aurora A-knockdown cells that expressed either empty vector or RNAi resistant Aurora A were measured on the indicated days. Data represent the means ± SD of three independent experiments.