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. 2021 Oct 11;10:e72833. doi: 10.7554/eLife.72833

Figure 4. Glc7-Bud14 interaction is required for spindle position checkpoint (SPOC) regulatory function of Bud14.

(A) Serial dilutions of indicated strains bearing URA3-based empty plasmid (not indicated on figure) or BUD14-containing URA3-based plasmids (BUD14 and bud14-F379A) were spotted on SC-URA plate and grown at indicated temperatures. (B) SPOC deficiency indexes of indicated strains carrying URA3-based empty plasmid (empty) or BUD14-containing URA3-based plasmids (BUD14 and bud14-F379A). Graphs are average of three independent experiments. A minimum of 100 cells were counted from each strain in each experiment. Error bars show standard deviation. *p<0.05 according to Student’s t-test. (C) Serial dilutions of indicated strains were spotted on 5-fluoroorotic acid (5-FOA) plate and grown at indicated temperatures. Cells that contain LTE1 on a URA3-based plasmid are indicated with pRS316-LTE1. (D) SPOC deficiency indexes of indicated strains. *p<0.05 according to Student’s t-test.

Figure 4—source data 1. Numerical data and statistics for Figure 4B.
Figure 4—source data 2. Numerical data and statistics for Figure 4D.

Figure 4.

Figure 4—figure supplement 1. Assessment of glc7-12 inactivation at different temperatures.

Figure 4—figure supplement 1.

(A) Serial dilutions of indicated strains were spotted on YPD and YPD-containing 0.01 M hydroxyurea (HU) and grown at indicated temperatures. The plates then were incubated at given temperatures for 2–3 days. (B) Analysis of cell morphology of GLC7 and glc7-12 cells at indicated temperatures. Liquid cultures were grown to log-phase at 23°C, shifted to indicated temperatures and further grown for 4 hr. Cells were fixed with ethanol and stained with DAPI. Cells were counted by microscopy and percentages of cells with large buds and single DAPI were plotted. A minimum of 100 cells were counted from each strain.