ALCAM in Melanoma Motility and Adhesion Figure 4. Expression of ALCAM in MUM-2C cells does not enhance wound-gap closure speed or invasive capacity. Immunostaining of MUM-2C cells (A) reveals that ALCAM expression is virtually undetectable in these cells. DAPI-stained nuclei are shown in blue; ALCAM staining is shown in red. In the 2C-ALC cell line (B), engineered to stably overexpress ALCAM, DAPI-stained nuclei are shown in blue, and ALCAM staining is shown in red. ALCAM localizes to points of contact between cells in the 2C-ALC cell line. The expression of ALCAM in 2C-ALC is confirmed by western blot in panel (C), and is undetectable in MUM-2C. Tubulin is shown as a loading control. The expression of ALCAM in 2C-ALC did not alter closure rate in a wound-gap assay (D), nor did it enhance invasive capacity of 2C-ALC cells when compared with MUM-2C in a transwell migration assay (t-test; PLoS ONE | www.plosone.org 8 June 2012 | Volume 7 | Issue 6 | e39330