Mutations in PLS affect cell process growth. (A) p190A^−/− MEFs expressing the indicated HA-tagged p190A constructs were plated overnight on fibronectin, fixed, and stained for nucleus (Hoechst, blue), cortactin (red), focal adhesions (vinculin, gray), and HA (green). Arrowheads show cortactin-rich edges, and arrows show focal adhesions. (B) Quantification of focal adhesion area in cells transfected as described in A (n = 15 cells per condition). Inset represents the number of focal adhesions (FA) per cell in the same experiment. (C) Quantification of ruffling cell process length. Graph presents the mean distance between nucleus and ruffle edges in cells transfected as described in A (n = 15 cells per condition). (D) Transfilter chemokinesis of cells transfected as described in A. Results are expressed as a percentage of basal migration without chemoattractant (n > 100 cells). For all graphs, values are expressed as the mean ± SEM of three (B and C) or five (D) independent experiments. Statistical significance was calculated relative to p190AWT condition. *, P < 0.05; **, P < 0.01; ***, P < 0.001. NT, nontransfected cells.