Fig. 4

Constitutive over-expression of DKK3 reorganizes cellular extensions and cell spreading. a-c SW-13 (a), SW-Neo (b), and SW-DKK3 (c) cells were grown on glass cover-slips, fixed, stained, and photographed. SW-13 and SW-Neo cells show a predominance of filopodia (red arrowheads) around edges; SW-DKK3 shows more lobopodia (small green arcs), absence of lamellipodia (blue arc), and few filopodia around edges. While cells in a and b appear to be polarized with filopodia at leading edge and lamellipodia at lagging edge, SW-DKK3 cells (c) show evenly spread flat lobopodia with extensive spreading and absence of polarity. Photomicrographs are taken using light microscope at 400× magnification. d Average number of lamellipodia, filopodia, and lobopodia per cell calculated from manual counting of cell extensions. Twenty randomly taken (400× magnification) photomicrographs of SW-13, SW-Neo, and SW-DKK cells used for quantification. e One hundred thousand SW-13, SW-Neo, and SW-DKK3 cells/well of 6-well plates were allowed to grow overnight, detached at specified times, cells remaining attached were fixed, stained, and counted manually