Figure 3.—

Chromosome spreads were performed on wild-type (PKY3412) and cac1Δ hir1Δ (PKY3415) cells that express CSE4–myc12 driven by the GAL1 promoter. Cells were grown in galactose-containing media and switched to glucose-containing media. Samples were taken at 0, 1, and 2 hr after repression with glucose. Immunofluorescence was performed with a rabbit anti-Mif2 antibody to visualize centromeres and a mouse anti-myc antibody to detect Cse4–myc. DNA was stained with DAPI. To image Cse4–myc in cac1Δ hir1Δ cells, the camera exposure time was half as long as that for Cse4–myc images for other cell types.