MMP-2 Promotes avb3 Mediated Tumor Cell Migration to 60 min). (C) Full-length fibronectin (FN) and MMP-2–cleaved fibronectin (FN+MMP-2+APMA) were electrophoresed in a native gel and transferred to nitrocellulose. An adhesion assay was performed using 1.06107/ml A375 cells incubated on nitrocellulose for 4 h. The membrane was washed and fixed, and the bound cells were stained. (D) The cleaved fibronectin fragments that cells can bind to were retrieved from the native gel, then coated on 24-well flat-bottom plates. An adhesion assay was performed using human A375 melanoma cells incubated in the coated plates for 30 min (*P,0.05). (E) An adhesion assay was performed using human M21 and M21-L melanoma cells incubated in the full-length fibronectin (FN)- or MMP- 2–cleaved fibronectin (FN+MMP-2+APMA)-coated plates for 30 min (***P,0.05). Statistical differences were determined by comparing the treated group with the normal control by t-test. doi:10.1371/journal.pone.0041591.g006 Figure S3 The effect of MMP-2 activity on the adhesion and migration of human DU145 prostatic cancer cells. (A) The effects of GM6001 on DU145 cell adhesion were analyzed in different concentration regarding its function on the different MMP individuals as described in MATERIALS AND METH- ODS. (B) DU145 cells were fluorescently stained and seeded on 48-well plates coated with 10 mg/ml human fibronectin (FN) or BSA as control. The cells were treated using RGD peptides or MMP-2I. The fluorescence intensity was measured using a fluo- rescence spetrophotometer. Statistical difference were determined by comparing treated group with the normal control by t-test (**, P,0.005). (C) The photomicrographs of DU145 cells in the agarose drop model 48 h after seeding. The arrows point the References 1. 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