Figure 3

Forced expression of TFF3 in MCF7 cells enhanced invasive phenotype. (A) Confocal microscopic visualization of the f-actin arrangement in MCF7 cells with either forced or depleted expression of TFF3. The red colour indicates f-actin, and blue colour indicates nuclei stained with DAPI. Images were captured under oil immersion X600 magnification. (B) Distribution of compact, loose, and scattered colonies of MCF7 cells with either forced or depleted expression of TFF3 as described in Methods. Left side, representative images of compact, loose, and scattered monolayer adherent colonies of MCF7, as observed with either forced or depleted expression of TFF3 in MCF7 cells. (C) Capacity of MCF7 cells with either forced or depleted expression of TFF3 to adhere to a Collagen I matrix. (D) Morphology of MCF7 cells with either forced or depleted expression of TFF3 when cultured on a Collagen I matrix. (E). Capacity of MCF7 cells with either forced or depleted expression of TFF3 to adhere to human microvascular endothelial cells (HMEC-1). The green colour indicates MCF7 cells (either forced or depleted expression of TFF3), and red colour indicates HMEC-1 cells. Images were captured under X100 magnification using a fluorescence microscope, as described in Methods. (F) Capacity of MCF7 cells with either forced or depleted expression of TFF3 to transmigrate through a HMEC-1 layer, as described in Methods. Statistical significance was assessed by using an unpaired two-tailed Student's t test (P <0.05 was considered as significant) using GraphPad Prism 5. Columns are the mean of triplicate experiments; bars, ± SD. ^**P <0.001, ^*P <0.05.