FIGURE 1:

Klp2 is required for nuclear clustering in SIN mutants, and its localization to MTs is modulated by the SIN. (A) klp2Δ sid2-250 and sid2-250 cells were grown to mid–log phase at 25°C and then shifted to 36°C for 2 h to inactivate Sid2. (Note that measurements of the distance between nuclei for both cell lines are shown in Figure 3A.) Cell outlines are marked with dashed lines. (B–E) Merged images show DAPI (blue), mCherry-Atb2 (α-tubulin; red), and Klp2-GFP (green). (B) sid2-250 mCherry-atb2 (tubulin) cells expressing Klp2-GFP were grown at 25°C and visualized after 90 min at 36°C. (C) Klp2-GFP was expressed in mCherry-atb2 cells. The selected field shows Klp2 localization during interphase (bottom and top cells) and during telophase (center). (D) Wild-type and sid2-250 cells expressing Klp2-GFP and mCherry-Atb2 were visualized during anaphase after 90 min at 36°C. (E) Klp2-GFP was visualized in cdc16-116 mCherry-atb2 cells after 2 h at 36°C. The scheme on the right represents the two different types of cell chambers generated in cdc16-116 cells after ectopic septation. In all cases, Klp2-GFP was expressed from the endogenous promoter. Cells were fixed with cold methanol. Images showing Klp2-GFP are shown inverted for clarity. DNA was stained with DAPI. Bar, 5 μm. All images are at the same magnification.