Fig. 1. pten^– cells exhibit fundamental defects in basic motile behavior and pseudopod formation in the absence of chemoattractant. Cells were distributed on the glass wall of a perfusion chamber at low density and perfused with buffer at a rate that turned over chamber volume four times a minute. (A,B) Perimeter and centroid tracks of representative parental AX2 (A) and pten^– (B) cells that had been pulsed for 6 hours with cAMP to achieve aggregation competence prior to motion analysis. (C,D) Perimeter and centroid tracks of representative AX2 (C) and pten^– (D) cells developed on pads to achieve aggregation competence prior to motion analysis. (E,F) Representative pad-developed parental AX2 (E) and pten^– (F) cell reconstructed in 3D with 3D-DIAS software over a 150 second or 125 second period, respectively, of cells migrating in buffer in a perfusion chamber. The arrows in A to D indicate net direction of cell migration. In E and F, pseudopods are colored red, the nucleus dark blue, tail fibers green and the cell body is shown in a transparent blue wireframe. Two views of the reconstructions are provided, at 30° and 90° angles. Note in F the abnormal extension of multiple lateral pseudopods by pten^– cells from the posterior as well as anterior end of the cell. See Movie 1 in supplementary material for a 3D dynamic presentation of an AX2 and pten^– cell migrating in buffer.