ADAMTS-12 and Trophoblast Invasion Figure 5. ADAMTS-12 regulates cell-extracellular matrix binding and invasion through an RGD-dependent mechanism. (A) Panels of photomicrographs of JEG-3 cells stably transfected with A12FL, A12Mut or LacZ. Cells were seeded onto 24-well plates pre-coated with ECM deposited by the same JEG-3 cell lines in a combinatorial fashion, and cultured for 30 min. Adjacent histograms show the number of cells that had bound to the pre-deposited native ECM, as described in Methods. (B) JEG-3 cells stably transfected with A12FL or LacZ were subjected to the native cell-ECM binding experiment described above, however cells were treated with either an RGD inhibiting peptide (RGD; 1 mM), a control peptide (RGDC; 1 mM) or untreated DMEM media (-). (C) A histogram representing the invasive indices of JEG-3 cells stably transfected with LacZ, A12FL or A12Mut. Cells were cultured in Matrigel-coated Transwell invasion chambers in complete DMEM media containing the RGD-inhibiting peptide (RGD; 1 mM), control peptide (RGDC; 1 mM) or complete DMEM media without treatment (-). Untreated cell lines were given an arbitrary invasion index of 1, and the invasive capacity of cells cultured in the presence of the control or RGD peptide were determined by generating an invasion index corresponding to their control cell line. All assays were performed in triplicate and performed on three independent occasions (n = 3; * = P#0.05, compared to controls). doi:10.1371/journal.pone.0018473.g005 PLoS ONE | www.plosone.org 9 April 2011 | Volume 6 | Issue 4 | e18473