FIGURE 3.

CDCP1-induced HeLa cell morphology change requires SFK activity. A, transmitted light microscopy analysis. HeLa vector, HeLa CDCP1, and HeLa CDCP1-Y734F cells were treated with vehicle (DMSO 0.075% v/v; control) or the SFK selective inhibitor SU6656 (10 μm) for 1 h. The media were then changed to normal growth media and after another 23 h cells were imaged using a Nikon Eclipse TE2000-U microscope. Arrow, epithelial morphology; arrowhead, elongated, fibroblastic morphology. Images are representative of three independent experiments. B, graph of the percentage of cells versus cell shape factor, determined using MetaMorph software analysis as described under “Experimental Procedures.” Black, HeLa vector cells; dark gray, HeLa CDCP1 cells; light gray, HeLa CDCP1-Y734F cells. C, graphic representation of flow cytometry analysis of cells after SU6656 treatment (10 μm 1 h) and re-equilibration for a further 23 h. Cells were lifted, and the percentage of cells undergoing apoptosis was assessed by flow cytometry analysis of cells stained with Alexa Fluor 647-conjugated annexin V. The data are from three experiments performed in triplicate. Black, HeLa vector cells; dark gray, HeLa CDCP1 cells; light gray, HeLa CDCP1-Y734F cells.