Cbl-b repressed IGF-I-induced EMT in gastric cancer cells. (A) Knockdown effect of Cbl-b was examined by Western blot. (B) The serum-starved cells were treated with or without IGF-I for 48 h. Photos were taken at × 20 magnification. (C) Immunofluorescent staining was performed as described above and visualized by fluorescence microscope at × 40 magnification. (D) Cell lysates were collected for Western blot analysis. (E) The migration assay was performed using the Boyden chamber methods as described in Materials and methods. Data are means ± SD in three independent experiments. * IGF-I untreated vs. IGF-I treated, p < 0.05; ** In IGF-I untreated group, ShRNA Cbl-b vs. NS Control, p < 0.05; *** In IGF-I treated group, ShRNA Cbl-b vs. NS Control, p < 0.05. E-cad, E-cadherin; Vim, Vimentin; ShRNA Cbl-b, Cbl-b shRNA transfected; NS Control, Non-silencing controls.