Figure 3.

Cx46-GFP increases the expression of EMT-related genes and EMT functional characteristics. (A) The EMT genes Snail, Zebt1, N-Caderin and Vimentin of MCF-7 and MCF-7Cx46-GFP were determined by qRT-PCR. The graph represents the means of three independent experiments. (B) In vitro scratch assays determined differences in breast cancer cell migration. The image in (B) was recorded every 2 h and photos were obtained at the indicated time points using a 10× objective on a Nikon inverted microscope, recorded and measured using the NIS-Element software. The figure shows images at 2 and 16 h for each cell line, and the color line shows the gap area in the three different cell lines evaluated. (C) Graphical representations of percentage of wound closure area of the three different experiments represented in B. The closing percentage was calculated at different times until the 16 h period was complete. Data represent the means of three independent experiments +/− SEM. * denotes p < 0.05, ** p < 0.01 and *** p < 0.001. (D) Representative images of fields in a Transwell migration assay of MCF-7 (up), MCF-7Cx46-GFP (center), and MDAMB-231 (bottom); cells that crossed the gel matrix were fixed and stained with Dapi. (E) Ten different fields were photographed, and the number of cells was counted and graphed. Data are presented as the means of three independent experiments +/− SEM.