Figure 6. The c-Src inhibitor collaborated with E₂ to promote EMT and increase cell migration.

(A) Changes of EMT biomarkers after different combinations treatment. Cell lysates of different treated cells were harvested. E-cadherin, N-cadherin, and fibrinogen were examined by immunoblotting. β-actin was detected for loading control. (B) Signaling pathways changes after different combinations treatment. Cell lysates of differently treated cells were harvested. Phosphorylated c-Src, MAPK, Akt were examined by immunoblotting. Total c-Src, MAPK, and Akt were detected for loading controls. (C) Transcription factors, Twist1 and Snail, were up-regulated after long-term combination treatment. Cell lysates of different treated cells were harvested. Twist1 and Snail were examined by immunoblotting. β-actin was detected for loading control. (D) Migratory capacities of MCF-7:PF cells, compared with MCF-7:5C cells. MCF-7:5C cells and MCF-7:PF cells were loaded in Boyden chambers as in Materials and Methods. Images were taken under bright field illumination at (×10) magnification (Olympus). (E) MCF-7:PF cells had higher migratory capacities than MCF-7:5C cells. Migrated cells were stained as in Materials and Methods. Cell numbers were counted in at least four microscopic fields at (10×10) magnification. P<0.05, * compared with MCF-7:5C cells. All the data shown were representative of at least three separate experiments with similar results.