Research Paper Volume 13, Issue 1 pp 1032—1050

SPOCK1/SIX1axis promotes breast cancer progression by activating AKT/mTOR signaling

SPOCK1 activates the AKT/mTOR signaling pathway in BC cells. (A) Proteins level on AKT/mTOR pathway of indicated cells were assayed by western blotting. GAPDH was used as a loading control. (B) Stable BC cells were treated with LY 290042 or Rapamycin. Then indicated protein levels were assayed by western blotting. GAPDH was used as a loading control. (C–E) Cell viability was detected in SPOCK1-overexpressed cells after treatment with LY 290042 or Rapamycin by MTT assay (C), Edu staining (D) and colony formation (E) assays. (F) Cell cycle progression was assayed by flow-cytometry analysis after dealing with LY 290042 or Rapamycin. (G) Stable BC cells were treated with LY 290042 or Rapamycin. Then cell cycle related protein levels were assayed by western blotting. GAPDH was used as a loading control. (H–I) Cell motility and invasion capacities was detected in SPOCK1-overexpressed cells after treatment with rapamycin or LY294002. (J) Stable BC cells were treated with LY 290042 or Rapamycin. Then levels of EMT-related proteins were assayed by western blotting. GAPDH was used as a loading control. (*PPPP

Figure 4. SPOCK1 activates the AKT/mTOR signaling pathway in BC cells. (A) Proteins level on AKT/mTOR pathway of indicated cells were assayed by western blotting. GAPDH was used as a loading control. (B) Stable BC cells were treated with LY 290042 or Rapamycin. Then indicated protein levels were assayed by western blotting. GAPDH was used as a loading control. (CE) Cell viability was detected in SPOCK1-overexpressed cells after treatment with LY 290042 or Rapamycin by MTT assay (C), Edu staining (D) and colony formation (E) assays. (F) Cell cycle progression was assayed by flow-cytometry analysis after dealing with LY 290042 or Rapamycin. (G) Stable BC cells were treated with LY 290042 or Rapamycin. Then cell cycle related protein levels were assayed by western blotting. GAPDH was used as a loading control. (HI) Cell motility and invasion capacities was detected in SPOCK1-overexpressed cells after treatment with rapamycin or LY294002. (J) Stable BC cells were treated with LY 290042 or Rapamycin. Then levels of EMT-related proteins were assayed by western blotting. GAPDH was used as a loading control. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001).