Fig. 2. Ace2p affects cell morphology and septin ring organization. (A) Wild-type (KGY246) cells containing the pREP3X-ace2 (pKG3141) plasmid were incubated at 25°C for 22 hours in the absence of thiamine to induce production of Ace2p. Cells were fixed in ethanol and stained with DAPI to visualize nuclei. (B) mid2-GFP (KGY2419) cells alone (left panels) or containing the pREP3X-ace2 (pKG3141) plasmid (right panels) were grown and visualized as in (A). (C) spn3-GFP (KGY3337) cells alone (top panel) or containing the pREP3X-ace2 (pKG3141) plasmid (bottom panel) were grown at 25°C for 22 hours in the absence of thiamine to induce Ace2p production. Cells fixed in 70% ethanol were visualized. (D) The spn3-GFP (KGY3337) and spn3-GFP ace2 (KGY4774) strains were grown at 25°C. Confocal images of live cells were obtained. Z-series optical sections were taken at 0.5 µM spacing. Images were rotated on the Z-axis to visualize the septin rings. Bars, 5 µm. Arrows indicate split ring structures that form after septation.