Extended Data Fig. 8: Segmentation of FAs and measurement of their area and average intensities of talin and vinculin in them.

(a) Segmentation and tracking of FAs was carried out using the FAA server [71]. The tracked FAs were then filtered to remove tracks with lifetimes of less than 5 minutes and tracks in which the size of FAs exceeded 0.5 μm² in more than 3 of the first 5 frames in order to remove tracks in which FAs were already mature when detected and leave only tracks corresponding to NAs. (b) After segmentation and tracking, all NAs were divided into two main groups - peripheral NAs and central NAs - using an eroded cell mask as described in Methods in the main text. Only peripheral NAs were used for comparison with the model. (c) The mean area of peripheral NAs measured in 4 different NIH-3T3 cells. (d, e) The mean normalized signal intensities of talin and vinculin measured in peripheral NAs in 4 different NIH-3T3 cells. The signal measured in each NA was normalized to the first frame, after which the mean normalized intensity was calculated by averaging over all peripheral NAs for each cell. In panels (c-e), the total number of tracked peripheral NAs in each cell was N = 133 in cell 1, N = 100 in cell 2, N = 101 in cell 3 and N = 103 in cell 4. The error bars in panels (c-e) represent SEM.