Figure 6. SIX1 involved in SPOCK1-mediated BC progression. (A) MDA-MB-231 and HS 578T cell line were transduced with si-con, si-SIX1#1, si-SIX1#2 and si-SIX1#3. The SIX1 levels in these were verified by western blot analysis after 48 h transfection. (B, C) Cell viability was detected in SPOCK1-overexpressed cells after transduction with si-RNAs by MTT assay (B) and colony formation (C) assay. (D) Cell cycle progression was assayed by flow-cytometry analysis after dealing with si-RNAs. (E) Stable BC cells were treated with si-RNAs. Then cell cycle related protein levels were assayed by western blotting. GAPDH was used as a loading control. (F, G) Cell motility and invasion capacities was detected in SPOCK1-overexpressed cells after treatment with si-RNAs. (H, I) Stable BC cells were treated with si-RNAs. The levels of EMT-related proteins and AKT/mTOR pathway were assayed by western blotting, respectively. GAPDH was used as a loading control. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001).