Figure 3

Knockdown of SNAT1 induces cell growth inhibition, cell cycle arrested, and apoptosis of breast cancer cells by inhibiting phosphorylation of Akt. (A) Western blot analysis of SNAT1 and p-Akt expression at 48 hours after the transfection of SNAT1-shRNA in 4T1 cells; (B) SNAT1-shRNA inhibits EGF induces p-Akt phosphorylation. 4T1 cells were transfected with SNAT1-shRNA with or without the presence of 50 ng/ml EGF. (C) 4T1 cells were transfected with SNAT1-shRNA or scrambled shRNA-transfected cells (sc-shRNA) at indicated times (24, 48, and 72 hours) and cell proliferation assay was performed; (D) 4T1 cells transfected with SNAT1-shRNA and sc-shRNA were grown in 6-well plates were incubated for 2 weeks. The numbers of the cell colonies were obtained and counted by 1-D gel quantity software QUANTITY ONE; (E) Cells were transfected with SNAT1-shRNA and sc-shRNA for 48 h. Then the cells were collected and the apoptosis rates were detected by flow cytometry; (F) After transfection for 48 h, 4T1 cells were harvested and cell cycle distributions were analyzed by flow cytometry. *P<0.05, ***P<0.001 considered statistically significant compared with sc-shRNA group.