FIGURE 3:

WRAMP structures in HT1080 cells. (A) Wide-field imaging (60×) of endogenous MCAM, F-actin, and myosin IIB in fixed HT1080 cells. Scale bars, 10 µm. (B) Cells treated without or with Wnt5a (250 ng/ml, 30 min) showing percentages of all cells with polarized MCAM, F-actin polarized along with MCAM, myosin IIB polarized along with MCAM, and F-actin plus myosin IIB polarized along with MCAM. Values show means and SEM from four or more slides (>800 cells) representing two or more biological replicates. *p < 0.05. The p values were calculated using standard two-tailed Student’s t test. (C) Live-cell imaging of HT1080 cells cotransfected with MCAM-GFP plus mCherry (also see Supplemental Movie S8) shows formation of WRAMP structures, followed by membrane retraction, not artifacts of cell volume or membrane thickness. Scale bars, 10 μm; times in hours:minutes.