Growth rate analysis does not depend on determining the complete traces of CCSs. (A, left) Thumbnails represent 12 temporal sections (24 s each) of a 288-s-long fluorescence movie of a BSC-1 cell stably expressing AP2-GFP. (right) CCS growth rate distribution of the entire movie. (B, left) Integrated movie segments (IMSs) are the quadrature sum of four 24-s-long segments in which density of CCSs is approximately four times larger than the original movie. Temporal separation is maximized between the segments to minimize the number of self-overlapping clathrin spots. (right) Growth rate distributions obtained from each IMS is plotted in comparison with the distribution obtained from the entire movie (black). (C) Growth rate distributions of the IMSs and the entire movie are displayed as bar plots. (D) We tested the reproducibility of growth rate distributions using the CCS traces produced by cmeAnalysis (Aguet et al., 2013) and TraCKer (homemade particle tracking program; see online supplemental software). Changes in slope distributions upon micropipette aspiration can be observed in both datasets. Numbers of the analyzed IMSs are 25 and 64 for before and during aspiration conditions, respectively (Ncells = 7). Error bars show standard deviations. FD, fast dissolution; FF, fast formation; P, plateau; SD, slow dissolution; SF, slow formation.
