ALCAM in Melanoma Motility and Adhesion Figure 7. ALCAM expression in 2C-ALC cells enhances formation of N-cadherin and ß-catenin junctions. (A, C) Immunostaining of 2C cells for nuclei (DAPI; blue), ß-catenin or N-Cadherin (green), and ALCAM (red) reveal that ß-catenin is diffusely localized and not prominent at cell-cell contacts. In contrast, 2C-ALC cells (B,D) display enhanced localization of ß-catenin and N-Cadherin to cell-cell contacts in the presence of ALCAM expression. In the merged images, ALCAM colocalizes at adherens junctions and at points of contact between adjacent cells. Levels of adhesion molecules were assayed in MUM-2C and 2C-ALC by western blot (E). The levels of pan-cadherin, N-cadherin, and ß-catenin are similar in both cell lines. The proportion of cells forming strong ß-catenin positive junctions is shown in F. 2C-ALC cells have significantly more ß-catenin positive cell junctions than do MUM-2C cells (t-test; p,0.05). Error bars are mean 6 S.E.M. Scale bars are 10 microns. doi:10.1371/journal.pone.0039330.g007 PLoS ONE | www.plosone.org 13 June 2012 | Volume 7 | Issue 6 | e39330