File:Plasticity-in-the-Macromolecular-Scale-Causal-Networks-of-Cell-Migration-pone.0090593.s008.ogv

English: Cell migration following ROCK-inhibition. ROCK-inhibitor-treated H1299-P/L cells expressing EGFP-Paxillin (green, left panel) and RubyRed-LifeAct (red, left panel) were imaged at 5 min intervals for 8 h. Note the reduced size and stability of EGFP-Paxillin-labeled CMACs, and the increased mobility and protrusiveness of this representative cell. After raw confocal image acquisition (left panel), automated image analysis facilitated segmentation of individual cells based on RubyRed-LifeAct fluorescence (cell boundary indicated by dark blue outline, overlaying EGFP-Paxillin channel, center panel). Subsequently, CMACs within each cell were segmented (CMACs detected based on EGFP-Paxillin fluorescence, indicated by red outlines, center panel). Finally, the centroids of both cells and their constituent CMACs were tracked via nearest neighbor analysis to facilitate time-resolved analyses of dynamic migratory processes (right panel). CMAC trajectories are color-coded for time, ≤10 time points shown. Scale bars in 1st frame = 50 µm.