Tumor-associated 3D ECMs induce relative organized MDA-MB-231 track orientations regulated by beta1-integrin but not by PI3K pathways. Time-lapse assays were carried out using MDA-MB-231 cells cultured on 2D or within staged 3D ECMs in the presence and/or absence of 10 and 50 nM Wortmannin and/or 50 μg/ml mAb13. Representative trajectory tracks were attained using Microsoft Excel as described in Methods. Orientation angles of the trajectories relative to the X-axis were rounded to the nearest 20th degree in order to identify a mode orientation angle. This mode angle was arbitrarily set as 0° and the original angles were rotated accordingly. Next, a new rounding of the rotated but otherwise intact data was performed only this time it was approximated to the 10th degree (see Methods for additional details). The percentages of angles positioned within 20° variance from the mode angle are indicated in the figure. Note that tumor-associated 3D matrices support oriented cell invasive trajectories (greater than 70% organization) in a beta1-integrin but not PI3K dependent manner.