Figure 3. Rate of pericentromere... | Rockefeller University Press

Figure 3.

$Figure 3. Rate of pericentromere chromatin stretching and recompaction. Time-lapse microscopy of sister pericentric CEN15 LacO arrays in mcm21Δ cells. Images were taken every 5 s for 200 s. Images were deconvolved as described in the Materials and methods, to determine the length of a LacO array along the spindle axis. (A–C) A representative time lapse shows stretching (line) and compaction (foci) of the LacO array over time. (A) Change in length of the left sister LacO array as a function of time in a single cell. The left LacO array appears predominantly as a spot and is slightly larger than the diameter of a diffraction spot. (B) Length of the right sister LacO array as a function of time. To determine a mean compaction and stretching rate, we fit a linear slope to regions that displayed greater than three successive steps in one direction. The mean compaction and stretching rates are −15 ± 12 nm/s (n = 9) and 13 ± 13 nm/s (n = 9), respectively, for six cells. (C) Selective images from the time lapse of sister LacOs A and B. The time point is indicated to the right. (D) Example from a time lapse indicating that sister chromatid stretching can switch from side to side. The right LacO array condenses as the left LacO array commences stretching. (E) Selective images from D. The time point is indicated to the right. Bars, 1 µm.$

Rate of pericentromere chromatin stretching and recompaction. Time-lapse microscopy of sister pericentric CEN15 LacO arrays in mcm21Δ cells. Images were taken every 5 s for 200 s. Images were deconvolved as described in the Materials and methods, to determine the length of a LacO array along the spindle axis. (A–C) A representative time lapse shows stretching (line) and compaction (foci) of the LacO array over time. (A) Change in length of the left sister LacO array as a function of time in a single cell. The left LacO array appears predominantly as a spot and is slightly larger than the diameter of a diffraction spot. (B) Length of the right sister LacO array as a function of time. To determine a mean compaction and stretching rate, we fit a linear slope to regions that displayed greater than three successive steps in one direction. The mean compaction and stretching rates are −15 ± 12 nm/s (n = 9) and 13 ± 13 nm/s (n = 9), respectively, for six cells. (C) Selective images from the time lapse of sister LacOs A and B. The time point is indicated to the right. (D) Example from a time lapse indicating that sister chromatid stretching can switch from side to side. The right LacO array condenses as the left LacO array commences stretching. (E) Selective images from D. The time point is indicated to the right. Bars, 1 µm.

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