Fig. 1

NESH/Abi-3 expression impaired WAVE2 membrane translocation and cell spreading. (a) Control NIH3T3 cells, and NIH3T3 cells expressing FLAG-Abi-1, FLAG-Abi-2, or FLAG-NESH were plated onto FN-coated coverslips. At the indicated times, the cells were fixed and stained with TRITC-phalloidin. (b) Quantitative analysis of cells in (a). Cells with lamellipodial structures were counted under a fluorescence microscope. At least 100 cells were analyzed for each sample. Data represent the means ± S.D. for three independent experiments. Error bars represent S.D. ***, P < 0.001, Student's t test. (c) The control and FLAG-NESH-expressing NIH3T3 cells were plated onto FN-coated coverslips. One hour after plating, the cells were fixed and stained with an anti-FLAG antibody, TRITC-phalloidin, an anti-WAVE2, or an anti-Arp3 antibody. The right panels show merged views of the TRITC-phalloidin and WAVE2 or Arp3 stained images. (d) The control and FLAG-NESH-expressing NIH3T3 cells were plated onto FN-coated coverslips. After overnight incubation, the cells were fixed and stained with TRITC-phalloidin. (e) The wound-healing assay was performed, and the migration area was quantified as described under Materials and Methods. Data represent the means ± S.D. for three independent experiments. The value for the control cells was set to 100 %. Error bars represent S.D. *, P < 0.05; n.s., the difference was not significant, Student's t test