Figure 4.

Experimental validation for the effect of selected mutations on metastasis. (a) Relative percentage of wound recovery. Scratch wound-healing assay was performed using MDA-MB-231 cells expressing the wild-type (WT) and mutant (MT) proteins of 13 selected genes. The relative recovery of wound region was calculated by cell confluency and is presented as bar graph. ***P<0.001; **P<0.01; and *P<0.05. (b) Relative percentage of migratory cells penetrating into the surrounding space. Transwell migration assay was carried out with MDA-MB-231 cells expressing the WT and MT proteins of eight selected genes. The representative staining images of migratory cells (upper) and the relative percentage values (lower) are shown, respectively. ***P<0.001; and **P<0.01. (c) EMT markers affected by the WT and MT protein expression. MDA-MB-231 cells were transfected with each plasmid encoding the WT and MT proteins, and the cell lysates were collected and subjected to the sodium dodecyl sulfate–polyacrylamide gel electrophoresis and immunoblotting. The gels have been run under the same experimental conditions.