Comparison between kymograph analysis and SkyPad.

A: First frame of a movie showing a myotube (white outline) with GFP-H1 nuclei (green). The red rectangle corresponds to the region used to create the kymograph in panel b. B: Kymograph showing nuclei moving in a myotube. Red arrows indicate the position where the nucleus moved at least 10 um from the previous arrow as calculated through SkyPad. White bar indicates a pausing or oscillating period, black bar indicates a moving period. Time in HH:min C: Speeds obtained by kymograph analysis and SkyPad between indicated frames. D: Trajectory of the nucleus represented with the coordinates in X and Y at each time point. The cross and gray circle represent the 10 um distance threshold used in SkyPad. The circle is traced on the first and last point of the trajectory (s and e respectively). Red dots correspond to arrow pointed positions in panel b. E, F: Speeds (e) and percentage of time in motion (f) of 18 nuclei were analyzed using kymograph and skypad analysis. Error bars correspond to S.E.M. Scale bars in a and b: 10 um.