Figure 5: Mob1 and Cdc15 localization during spindle misalignment.

(a–c) Representative still images from the time-lapse series of MOB1-GFP mCherry-TUB1 kar9Δ cells with misaligned spindles in the presence of KIN4 and SPO12 (a), in the absence of KIN4 (b) and in the absence of KIN4 and SPO12 (c). In (a), cell 1 has a normal aligned spindle, whereas cell 2 has a misaligned spindle. Arrows indicate the SPB or bud neck localized Mob1 when detectible. Scale bars: 3 μm. (d) Analysis of Mob1-GFP SPB binding during spindle misalignment. Time-lapse series of 30 cells were analyzed per strain. The graph shows the percentage of cells with misaligned spindles in which Mob1-GFP remained low or accumulated at SPBs during anaphase. (e) Box-and-whisker plots showing the mean fluorescence signal intensities of Mob1-GFP and Cdc15-GFP at SPBs of cells with misaligned spindles. The boxes show the lower and upper quartiles, the whiskers show the minimum and maximal values excluding outliers; outliers (shown as dots) were calculated as values greater or lower than 1.5 times the interquartile range; the line inside the box indicates the median. Asterisks mark significant differences according to two-tailed Student’s t-test (P<0.01). For Mob1-GFP plot, 100 SPBs were quantified per strain. For Cdc15-GFP plot, 50 signals were quantified per strain. (f) Percentage of SPOC-deficient phenotypes in the indicated cell types. Graphs show an average of three independent experiments. Error bars show s.d. Per experiment, 100 cells were counted per cell type. Asterisk indicates significant differences according to two-tailed Student’s t-test (P<0.05).