Figure 2: hSpry1 overexpression inhibits uPAR-stimulated cell migration. (a) HEK293/uPAR, (b) HCT-116, and (c) MDA-MB-231 cells were transfected with pCDNA3.1 (light bars) or pCDNA3.1/hSpry1 (dark bars) and examined using wound healing assay. (d) A monolayer of confluent cells (MDA-MB-231 cells in these photos) was wounded with a 200 μL pipette tip, and the closure of the “scratch” was observed in the presence of either 10% FCS, 1% FCS, or 100 ng/mL of EGF, uPA, or ATF. Photographs taken immediately after the creation of a “scratch” are marked as 0 h. Photographs taken 18 h later demonstrate the differential extent of migration for the various cell lines. Cells migration was assessed by measuring the distance of the wounded region lacking cells in pixels cells after 18 h. Bars are mean ± SEM ( ), and significance was measured using unpaired t-test ( ; ). Data are representative of 3 independent experiments.