Figure 2.

N-cadherin could function as a receptor for CS-E and enhanced the invasive activity of BT-549 cells. A, cells pre-treated with (+) or without (−) anti-N-cadherin antibody were added to the CS-E-coated wells, and cells bound to the CS-E on the plate were estimated using cellular lactate dehydrogenase activity as an indicator (n = 3). Statistical significance was assessed using a Student's t test. B, TCF reporter activity in the absence (−) or presence (+) of anti-N-cadherin antibody and/or CS-E was measured (n = 3). C, BT-549 cells cultured in a monolayer were scratched and subsequently cultured for the indicated periods in the presence of CS-E, anti-N-cadherin antibody, or both. Dashed lines show the edge of the gap newly created by scratching. D, invasive activity of BT-549 cells was measured in the absence or presence of CS-E and/or anti-N-cadherin antibody (Ab) (n = 3). E, BT-549 cells were stimulated with CS-E untreated (No treated) or pre-treated at 94 °C for 20 min (Heat-treated), and their invasive activities were measured (n = 3). F, invasive activities of BT-549 cells digested with chondroitinase ABC or pre-treated with heparin (50 μg/ml) were measured (n = 3). G, either EGFP- or N-Cad(Full)-EGFP-expressing MCF7 cells were treated with or without CS-E, and their invasive activities were measured (n = 3). H, panel a, cell lysates of BT-549 and HC1954 cells were analyzed by immunoblotting using an anti-N-cadherin antibody. Panel b, β-catenin transcriptional activities of HCC1954 cells in the presence or absence of CS-E were measured using TCF reporter vector (n = 3).