Fig. 5

Single molecule tracking of 1 × HA-tagged proteins in cells. a The mean positions of tracks of single frankenbody (FB) bound to 1 × HA-H2B provides a mobility map of H2B across the cell nucleus (10,949 tracks were generated from 977,516 total FB localizations). Tracks are color coded according to their average frame-to-frame jump size. The lighter colored tracks with relatively small jump sizes are enriched along the edge of the cell nucleus, where heterochromatin is typically enriched. To ensure tracks represent FB bound to HA-H2B, tracks were filtered such that their length had to be at least 16 consecutive frames and jumps between frames had to all be <220 nm. Full tracks within the yellow box are displayed in the zoom-in on the right. See also Supplementary Movie 2. b In cells expressing HA-H2B, the number of filtered FB tracks were between one and two orders of magnitude greater than in control cells lacking HA-H2B, demonstrating false-positive tracks are rare (see Supplementary Fig. 5). c The average mean-squared displacement of tracks provides a good estimate for average HA-H2B mobility. All tracks are from n = 9 cells in three independent experiments. Student’s t-test. Error bar, cell-to-cell SD of the average MSD. Scale bar, 5 µm. Source data are provided as a Source Data file. For the box and whisker plots, median is shown by a white line, the box indicates 25–75% range, and whiskers indicate 5–95% range