Depletion of km23-1 inhibits cell migration and invasion of human CRC cells.

<p><b>A:</b> Transwell migration assays were performed as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#s2" target="_blank">Materials and methods</a>.” Briefly, HCT116 cells stably transduced with lentiviral vectors expressing either piLenti-NC siRNA-GFP or piLenti-km23-1 siRNA-GFP were seeded into the upper wells of the Costar Transwell System (8-µm pore size polycarbonate membrane, 6.5-mm diameter), and the cells on the lower surface of the well after 24 h were fixed in methanol and stained with DAPI. <b>Top</b>, representative images of the lower surface of the membrane are shown (100× magnification). <b>Bottom</b>, the number of migrating cells of both NC siRNA- and km23-1-siRNA-HCT116 stable transfectants were counted under a fluorescence microscope and statistically analyzed. *p<0.01 compared to NC siRNA. <b>B:</b> Matrigel invasion assays were performed with the indicated RKO stable cells clones (clones #1, 5) for 24 h using EGF (20 ng/ml) as the stimulus as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#s2" target="_blank">Materials and methods</a>.” Invaded cells were stained with 0.2% crystal violet. <b>Top</b>, representative images of the lower membrane surface from Matrigel are shown (100× magnification). <b>Bottom</b>, the number of invading cells for both NC siRNA and km23-1-siRNA HCT116 stable transfectants were counted under a light microscope and statistically analyzed. *p<0.01 compared to EV.