Persistence and speed of cell migration regulate migrasomes formation in MGC803 and NRK cells. (A and E) Representative images from a time-lapse video of WT MGC803 cell (A) and NRK cell (E). White arrows indicate the migration direction, red circles indicate migrasomes, and white boxes indicate ROIs of T-region and P-region. Scale bar, 20 µm. (B and F) Analysis of RFs and migrasomes in T-region and P-region depicted in A and E, respectively. Images were inverted, and migrasomes and RFs were masked on the right. n in middle panel represents the number of migrasomes. Scale bars, 10 µm. (C and G) Quantification of the number of parental RFs (green lines) and the ratio of rear end width to cell perimeter (red lines) of the migrating cell in A and E, respectively. (D and H) Representative images from time-lapse videos of WT MGC803 cells (D) and NRK cells (H). The left panel shows an example of a slow persistent migrating cell, and the right panel shows an example of a fast persistent migrating cell. The red and blue lines indicate the start and end position of the migrating cell, respectively; yellow arrows indicated the position of migrasome formation; and black outline arrows indicated the displacement of the migrating cell. Scale bar, 20 µm.