Fig. 5

ESPN promotes OS cells migration and invasion through PI3K/AKT/mTOR/ZEB1 pathway (A) Western blot analysis of phosphorylated and total PI3K, AKT, and mTOR protein expression in control and ESPN knockout OS cells. (B) The schematic diagram illustrates the binding interaction between ESPN and PI3K, with PI3K depicted in brown and ESPN represented in yellow. (C) Immunoprecipitation and Western blot analysis demonstrated the interaction between MYC-tagged PI3K and HA-tagged ESPN in 143B and MNNG-HOS cells. (D) The protein expression levels of mTOR, p-mTOR, and ZEB1 were detected in different groups (sh-CON, sh-1 and sh-1 + 3BDO) of 143B and MNNG-HOS cells. (E) The migration and invasion abilities of 143B and MNNG-HOS cells in different groups (sh-CON, sh-1 and sh-1 + 3BDO) were assessed using transwell and invasion assays. (F) Determination of PI3K, AKT and mTOR phosphorylated proteins and total protein expression before and after ESPN overexpression in OS cells by Western blot. (G) The protein expression levels of mTOR, p-mTOR, and ZEB1 were measured in different groups (Vector, ESPN and ESPN + Rapamycin) of MG63 cells. (H, I) The migration and invasion abilities of MG63 in different groups (Vector, ESPN and ESPN + Rapamycin) were assessed using transwell and invasion assays. These data represent three independent experiments. Scale bar = 50 μm