Effect of cytochalasin D treatment on ECM patterning of endogenous TSP proteins expressed by SW480 and RCS cells
(A) Representative bright-field images and confocal XY sections of SW480 and RCS cells treated with either 0.25 μM (SW480) or 1 μM (RCS) cytochalasin D or DMSO solvent only for 18 h. For confocal microscopy, cells were stained with FITC–Phalloidin (green) and DAPI (blue) prior to imaging. (B) Representative fluorescence images of ECM deposition patterns of endogenous TSP1 by SW480 cells, after treatment of cells with 0.25 μM cytochalasin D or vehicle (DMSO) for 18 h. An image of SW480 ECM without primary antibody is included as a control. (C) Quantification of edge concentrated ECM puncta of endogenous TSP1 by SW480 cells,±cytochalasin D. (D) Representative fluorescence images of ECM deposition patterns of endogenous TSP5 by RCS cells, after treatment of cells with 1 μM cytochalasin D or vehicle (DMSO) for 24 h. For cytochalasin D treated cells, three separate images are shown at the same scale and arrowheads indicate circular ECM arrays. An image of RCS ECM without primary antibody is included as a control. In (C), columns represent the mean and bars indicate S.E.M. from four independent experiments. *P<0.05.