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Fig. 2. Dynein-dependent nuclear rotations are enhanced after monolayer wounding. (A) A time series of a nucleus that rotates during cell migration. Red arrows indicate two nucleoli that stay in position relative to other nucleoli within the nucleus. This nucleus rotates counter-clockwise for the first 30 minutes, and then switches direction during the final 10 minutes of this timelapse. Scale bar, 10 µm. Time is minutes:seconds. (B) Tracks of nucleoli during migration of fibroblasts over 45 minutes. Large circular traces seen in mock-transfected cells demonstrate paths of nucleoli in nuclei that are rotating. These are rarely seen in DHC and p150^Glued siRNA cells (n=15). Axis labels are in µm. (C) Particle tracking was used to track the duration of rotation of nuclei from cells at the wound edge during migration. A run was defined as rotation in a single direction without pausing (n=225 runs from 25 cells). (D) Angular velocity of nuclei in cells along the wound edge of wounded cultures is higher than that of nuclei in cells that have been grown to confluency, but not wounded (n=25). (E) Angular velocity of nuclei in mock-treated cells along the leading edge is greater than that of DHC and p150^Glued siRNA cells, which rarely rotate and thus have a slower average angular velocity (n=15). Error bars indicate s.e.m.; ^*P<0.05.

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