Figure 6
AGK downregulates FOXO1 transaactivity and activates the AKT pathway. (A) Relative FOXO1 reporter activity in AGK-infected cells, AGK-shRNA infected cells and control cells. (B) Western blotting analysis of phosphorylated-Akt (p-Akt), total Akt, phosphorylated-GSK-3β (p-GSK-3β), total GSK-3β, phosphorylated-FOXO1 (Ser256) and total FOXO1 proteins in MGF-7 and SKBR3 breast cancer cells. GAPDH was used as a loading control. (C) Expression (upper panel) and correlation analysis (lower panel) of AGK and FOXO1 expression in 10 freshly isolated human breast cancer samples. (D) Western blotting analysis of p21Cip1, p27Kip1, cyclin D1 and FOXO1 proteins in the indicated breast cancer cell lines. GAPDH was used as a loading control. (E, F) The MTT assay shows that silencing FOXO1 increased the proliferation of AGK-silenced cells as determined by an MTT assay. Error bars represent SD from three independent experiments. *P < 0.05.