Figure 3.

Mdp3 stimulates the motility of breast cancer cells. (A) Immunoblot analysis of Mdp3 and β-actin expression in MDA-MB-231 and Hs578T cells transfected with control or Mdp3 siRNAs for 48 hours. (B) Control or Mdp3 siRNA-transfected cells were scratched, and wound margins were imaged 0 or 24 hours later. (C) Experiments were performed as in B, and the extent of wound closure was analyzed. (D) Control or Mdp3 siRNA-transfected cells were plated into inserts precoated with matrigel, and cells underside the porous membrane were stained with crystal violet 24 hours later. (E) Experiments were performed as in D, and the extent of cell invasion was quantified. (F) Experiments were performed as in D, except that the inserts were precoated with fibronectin (FN), poly-D-lysine (PDL), or gelatin (GEL). (G) Movement tracks of MDA-MB-231 cells transfected with control or Mdp3 siRNAs. (H) Experiments were performed as in G, and the Euclidean distance and the accumulated distance were determined. (I) Experiments were performed as in panel G, and the velocity of cell movement was calculated. (J) Immunoblot analysis of Mdp3 and β-actin expression in MCF-10A and MDA-MB-231 cells transfected with control or Mdp3 for 48 hours. (K) Control or Mdp3-transfected cells were scratched, and the extent of wound closure was analyzed 24 hours later. (L) Control or Mdp3-transfected cells were plated into inserts precoated with matrigel, and the extent of cell invasion was quantified by crystal violet staining 24 hours later.