Fig. 4

FTL promoted EMT through AKT/GSK3β/β-catenin signaling. a Western blotting analysis of AKT, p-AKT (ser473),GSK3β,p-GSK3β(ser9), β-catenin and p-β-catenin was performed in FTL silenced or FTL forced U87 and U251 cells. GAPDH was used as loading control. b Cytoplasmic and nuclear fractions of U87and U251 cells were extracted separately, and then expression of β-catenin was detected by western blot. Tubulin as cytoplasmic loading control and Histone H3 as nuclear loading control. c-d Next, TCF/LEF luciferase reporter assay was used to detect activity of β-catenin signaling in FTL-shRNA or FTL-plasmid transfected glioma cells. ***, P < 0.001. e Immunefluorescence staining confirmed that the distribution of β-catenin inside the cell. DAPI was used for nuclear staining; Scale bars,10 μm. f-h U87 and U251 cells transfected with NC-shRNA or FTL-shRNA were treated with DMSO or IM-12(GSK3β inhibitor) for 24 h.Transwell assay was used for invasion detection. g Western blot was employed for measurement expression of vimentin,snail1 and β-catenin. Three independent experiments were performed. *,P < 0.5;**, P < 0.01;***, P < 0.001