Fig. 5. Spontaneous ruffling activity of stable CHO clones expressing EGFP-1-448 mDia1. (A) Immunoblots of control CHO cells (lane 1), CHO transfected with the EGFP-containing vector (lane 2), and two clones (termed 1A and 1B) stably transfected with EGFP-1-448mDia1 (lanes 3 and 4), as obtained with the polyclonal mDia1 antibody. Note that these clones express the fragment in amounts approximately equal to the endogenous mDia1. (B) Proportion of spontaneously ruffling cells in the mock-transfected (CHO/EGFP) and clone 1A cells. Cells were fixed and examined by Nomarski optics and for EGFP-fluorescence, and classified according to the presence of ruffles (black bars) or no ruffles (white bars). Approximately 300 cells were counted for each class, in each of 4 independent experiments.