Fig. 3. Phospho-defective WAVE2 expression does not affect ruffling in response to PDGF. (A) NIH 3T3 cells stably expressing EGFP, EGFP-WAVE2-wild-type (W2 WT) or EGFP-WAVE2-AAA (W2 AAA) were plated on fibronectin-coated glass coverslips overnight in 0.5% serum. PDGF was added where indicated for 25 minutes before cells were fixed, permeabilised and stained with phalloidin (F-actin). Bar, 30 µm. (B) Experiments were performed as for A, then six random microscopic fields were imaged and the number of cells with one or more ruffles were counted. Dark bars (+) represent cells with ruffles, and light-grey bars (–) represent those without ruffles. Statistical analysis with Fisher's exact test revealed no significant differences between cell lines in either starved or stimulated conditions.