Depletion of km23-1 reduces Ezrin expression in human CRC cells.

<p><b>A:</b> The cell lysates used for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#pone-0066439-g001" target="_blank">Fig. 1D</a> were further subjected to Western blot analysis of Ezrin expression <b>(Top). Bottom</b>, DIC loading. <b>B:</b> HCT116 cells were infected with either pilenti-NC siRNA-GFP or the pilenti-km23-1 siRNA-GFP set. 24 h after infection, Western blotting was performed using the indicated antibodies. <b>Top</b>, confirms knockdown of endogenous km23-1. <b>Bottom</b>, DIC protein was assessed as a loading control. <b>C:</b> HCT116 stable pools that migrated through to the lower membrane in the Matrigel invasion assay were stained with an Ezrin Ab, followed by cy3-conjugated goat anti-rabbit IgG (red). DAPI staining permitted visualization of nuclei of individual cells (blue). GFP was used as a marker to designate cells transduced with siRNA (green). The cells were analyzed by an Olympus IX81 microscope at a magnification of 1000× with the appropriate filter sets.