The effects of S100A14 and S100A16 knockdown on cancer cell invasion and motility. A, In vitro invasion assay using a Matrigel-coated transmembrane. Invading MCF7 and SK-BR-7 cells were counted 24 h after seeding of knockdown cells (the error bar represent S.D., n=3). B, Wound healing assay using MCF7 cells transfected with siRNA for S100A14or S100A16. The wound width was measured, and the distance over which the cells had migrated 48 h after scratching was calculated (the error bars represent S.D., n=3). C, A dual-color wound healing assay using MCF7 cells that were stably transfected with GFP or Tomato expression vectors that were further transfected with siRNA for S100A14 or control, respectively. The white lines represent the migration fronts at 48 h after scratching of a mixed culture of cells on a confluent monolayer. Scale bars; 50 μm. D, Confocal images of representative spheroids of MCF7 cells grown in a collagen gel for 48 h after transfection of S100A14 or control siRNA. Scale bar; 50 μm.