Fig. 2

GAS7b inhibits MDA-MB-231 breast cancer cell migration, invasion, cell adhesion, and FA formation. MDA-MB-231 cells were transfected with control or GAS7b-expressing plasmids. After 48 h, the cells were subjected to the following assays. a Upper: representative photographs of the trans-well cell migration and invasion assays. Bottom: statistical analysis of migrated and invaded cells are shown by histograms. Data represent normalized mean ± SD (n = 3). b Upper: representative photographs of wound healing cell migration assay. Bottom: statistical analysis of cell migrated areas are shown by histograms. Data represent normalized mean ± SD (n = 3). c Upper: representative photographs of the cell adhesion assay using fibronectin, type I collagen, or laminin coated plates. Bottom: the adhesion cells were stained with crystal violet, and the eluted dye was measured by spectrophotometer. Data represent normalized mean ± SD (n = 3). d Immunofluorescence staining and confocal microscopy of Vinculin (green) for FA, Phalloidin (red) for F-actin, and DAPI (blue) for nuclear staining. MDA-MB-231 cells were transfected with control vector, GAS7b-expressing plasmid, or GAS7b-expressing plasmid plus GAS7 siRNA, and cells were plated on fibronectin coated coverslips for 90 min, and e quantification of total focal adhesions per cell (n = 10, mean ± SD). Two-tailed t-test was used for these statistical analysis (*p < 0.05; **p < 0.01; ***p < 0.001)