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. 2016 Dec 30;205(3):1169–1178. doi: 10.1534/genetics.116.195727

Figure 4.

Figure 4

rts1Δ mutants are sensitive to CIK1-CC overexpression. (A) Overexpression of CIK1-CC is toxic to rts1Δ cells. WT and rts1Δ cells with a vector (V) or a PGALCIK1-CC (CC) plasmid were 10-fold serial diluted and spotted onto glucose and galactose plates for further incubation at 30° for 2 days. (B) rts1∆ cells lose viability after CIK1-CC overexpression. WT and rts1∆ cells with a vector or a PGALCIK1-CC plasmid were grown to log phase in raffinose medium at 30° and then galactose was added into the medium. Cells were collected at 0, 3, and 6 hr and spread onto YPD plates to assess the plating efficiency (n ≥ 200). (C) CIK1-CC overexpression leads to chromosome missegregation in rts1Δ cells. CEN4-GFP TUB1-mCherry and rts1Δ CEN4-GFP TUB1-mCherry cells with a vector or a PGALCIK1-CC plasmid were arrested in G1 phase in raffinose medium and then released into galactose medium at 30°. Cells were collected at indicated time points and fixed for the examination of fluorescence signals. The percentage of cells with missegregated sister CEN4-GFPs among all anaphase cells is shown on the top left (n > 100). The localization of CEN4-GFP as well as spindle morphology in some representative cells is shown at the bottom. The budding index is shown on the right. Bar, 5 µm.