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. 2016 Aug 15;30(16):1881–1894. doi: 10.1101/gad.282194.116

Figure 4.

Figure 4.

Overexpression of specific ChrI segments suppresses ulp2Δ aneuploidy. (A) Strategy used to identify specific regions of ChrI that suppress ulp2Δ aneuploidy. A ulp2Δ/YCplac33-ULP2 strain (MHY1379) was transformed with the indicated pGP564 (LEU2)-based ChrI library plasmids. Cells were struck on SD-Leu + FOA twice to evict YCplac33-ULP2. This strategy was used in subsequent ploidy assays as well. (B) Following YCplac33-ULP2 eviction, qPCR was used to assay chromosome copy number of ulp2Δ cells bearing the ChrI plasmids. Copy number was determined in relation to the ulp2Δ + ULP2 euploid strain. The ratio of ChrI to ChrIII copy number is plotted. (C,D) Schematic diagrams of plasmids pGP564-4 (C) and pGP564-10 (D). Locations of ORFs (gray arrows) and SpeI, HindIII, and NotI restriction sites are indicated. Subclones used in E and F are shown below the ChrI regions. “10-5” shows the ChrI segment that was inserted into plasmid pRS315-CCR4 and used in Figure 5, C and D. (E,F) qPCR ploidy assays of ulp2Δ cells with the high-copy plasmids shown in C and D. Chromosome copy number was determined as in Figure 1C. The error bars indicate the SD from three PCR reactions with two independent genomic DNA preparations.