FIGURE 8:

Persistence of Smc3 and Mcd1 close to spindle pole bodies in anaphase cells of cdc5-99 strains. (A) Wild-type (WT, KBY6098-1) and cdc5-99 (KBY6097-1) strains containing SMC3-GFP and SPC29-RFP were grown at permissive temperature (25°C) and shifted to the nonpermissive temperature (37°C) for 2.5 h. Cells with spindle length 3–8 μm were considered to be in anaphase and were used for imaging. Spc29-RFP was used as a spindle pole marker. Representative images showing the persistence of Smc3-GFP signals in anaphase cells of cdc5-99 strain (KBY6097-1). (B) Percentage of Smc3-GFP foci proximal to the spindle poles was determined in cdc5-99 strain (KBY6097-1) at permissive temperature (25°C, n = 125) and 2.5 h after shift to nonpermissive temperature (37°C, n = 144). Standard weighted-means analysis was used to calculate averages, and significance was determined by one-way analysis of variance. Bar diagram represents average ± SE of the mean. *p < 0.05. (C) WT (YMB9695) and cdc5-99 (YMB9696) strains containing MCD1-GFP and SPC29-RFP were used. Strain VG3507-2A carrying MCD1-GFP (a gift from Vincent Guacci, University of California, Berkeley, CA) was used to construct strains YMB9695 and YMB9696. Strains were grown at permissive temperature (25°C) and shifted to the nonpermissive temperature (37°C) for 3 h. Cells with spindle length 3–8 μm were considered to be in anaphase and were used for imaging. Spc29-RFP was used as a spindle pole marker. Representative images showing the persistence of Mcd1-GFP signals in anaphase cells of cdc5-99 strain (YMB9696). (D) Percentage of Mcd1-GFP foci proximal to the spindle poles was determined in cdc5-99 strain (YMB9696) at permissive temperature (25°C, n = 94) and 3 h after shift to nonpermissive temperature (37°C, n = 197). Standard weighted-means analysis was used to calculate averages, and significance was determined by one-way analysis of variance. Bar diagram represents average ± SE of the mean. *p < 0.05.