(A) Schematic showing SPB cycle, including the metaphase arrest following depletion of CDC20 in MET-CDC20 strains and the α-factor arrest point. The color of each satellite component used in the figure is noted. (B–D) Metaphase-arrested MET-CDC20 strains containing Spc42-mTurquoise2 and Nud1-YFP (SLJ9720, B), Nud1-mTurquoise2 and Spc42-YFP (SLJ9685, C), and Spc42-mTurquoise2 and Spc29-YFP (SLJ10009, D) were released into the cell cycle using SC-methionine media. Aliquots were taken at the indicated times for analysis of satellite assembly by SIM. Cells were also released into α-factor for the indicated time. (B, C) On the left, the percentage of SPBs that contain two foci Nud1 and 0, 1, or 2 overlapping foci Spc42 are plotted. On the right, the percentage of SPBs that contain two foci Spc42 and 0, 1, or 2 overlapping foci Nud1 are plotted. Note, B and C differ in the fluor; the fact that virtually identical results were obtained in both experiments suggests that mTurquoise2 and YFP have similar maturation rates and can be used to detect the newly forming satellite. (D) On the left, the percentage of SPBs that contain two foci Spc29 and 0, 1, or 2 overlapping foci Spc42 are plotted. On the right, the percentage of SPBs that contain two foci Spc42 and 0, 1, or 2 overlapping foci Spc29 are plotted. (E) Asynchronously grown cells containing Nud1-mTurquoise2 and Spc42-YFP (SLJ9275, left), Spc42-mTurquoise2 and Nud1-YFP (SLJ9660, center), Spc42-mTurquoise2 and Spc29-YFP (SLJ8082, center right), and Spc29-CFP and Spc42-YFP (DHY47-6B/SLJ10596, right) were imaged by SIM. The percentages of unbudded cells with a single SPB with a single focus of Spc42 and one or two foci of Nud1 or Spc29; or a single SPB with two foci of Spc42 and one or two foci of Nud1 or Spc29 was plotted. A small fraction of cells (2–7%) have other configurations that are not depicted, such as SPBs labeled with only one fluorescent protein. N > 77 in all samples. Related to Figure 5C–E.