Effect of QSOX1 on invasiveness. (A, B) Cells were seeded on polycarbonate filter coated with Matrigel, in the presence/or not of a specific inhibitor of MMP-2 (OA-Hy) at 10 μM and incubated for 24 h and analyzed as described in Materials and Methods. Inserts were stained with 2% crystal violet and photographed. * P <0.05, compared to control (Wilcoxon test). (C, D)ZY Gelatin Zymography of matrix-metalloproteinase in conditioned medium. Cells were seeded and the media were collected after 24 h and 12 h of serum starvation for the MCF-7 and MDA-MB-231 cell lines respectively. Conditioned medium from HT-1080 cell line was used as positive control. Vertical line has been inserted to indicate a repositioned gel lane. (C, D)WB Blotting identification of MMP-2 in protein extracts from conditioned culture medium. (E, F) Cells were seeded on polycarbonate filters coated without ECM and incubated for 24 h and analyzed as described in (A). * P <0.05, compared to control (Wilcoxon test). These experiments are representative of three separate assays.