FIGURE 5:

Effect of blocking WRAMP structures on cell speed and wound healing. To measure cell speed, (A) WM239a and (B) A375 cells were transfected with PB-H2B-mCherry and seeded on glass-bottom 96-well plates, followed by scratch wounding. Every cell that migrated into the wound was tracked using a cell-tracking algorithm, calculating movement speed for individual cells. Bars show the mean and SEM from three wells (>360 cells/condition for WM239a cells and >180 cells/condition for A375 cells). *p < 0.01 and **p < 0.005. The p values were calculated using standard two-tailed Student’s t test. (C–F) Scratch wound assays were used to measure of the effects of suppressing Wnt5a signaling and MCAM expression on wound healing. Cells were imaged using bright-field microscopy in plastic 96-well dishes, and four wells were measured for each condition at various time points. We quantified (C) WM239a and (D) A375 cells in the presence of 10% FBS and (E) WM239a and (F) A375 cells in the absence of serum. Two-way analysis of variance was used to determine the significance between the treatments and nontargeting siRNA control across the time points. ^‡Treatment is significantly different compared with nontargeting control (NTC) siRNA with p < 0.0001.