Fig. 5

Phosphorylation of ELMO1 Y724 contributes to the enhancement of cell adhesion. NIH3T3-Dock180-Control, −wild-type ELMO1, and - Y724F ELMO1 cells were employed throughout the analyses in Fig. 4. a Representative cell morphologies onto the conventional culture dish are shown. Black and white arrowheads indicate the apparent membrane ruffles and the protrusion of the cell membrane, respectively. b-d The trypsinized cells were replaced onto glass-based dishes coated with 10 μg/ml fibronectin. b Immediately after re-plating, DIC images of cells were obtained every one minute for 1 h using a time-lapse imaging system. The micrographs after 30 min are shown. To define the cell size, the margins of several cells are marked with dotted lines. c Cells at 30 min after re-plating were subjected to actin staining. d In immunofluorescence described in Fig. 5c and 5e, the sizes of the individual cells (left panel) and the numbers of paxillin (right panel) were determined using MetaMorph software and manually, respectively. The data are shown as mean ± SD of six cells from a single experiment. *, P < 0.001; **, P < 0.00001. e Cells at 30 min after re-plating were subjected to immunofluorescence analysis with Ab to paxillin and with phalloidin for actin staining. In D, representative micrographs are shown and the merged images placed at the right. f The cells were replaced onto the 96-well plates coated with fibronectin. After incubation for 90 min, the bound cells were stained, lysed, and measured as described in Materials and Methods. The data are shown as mean ± SD of 3 wells on each condition. *, P < 0.005; **, P < 0.0001