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Fig. 3. Mid2p functions in parallel with other Ace2p targets. (A) The spn3-GFP agn1{Delta} (KGY5091), spn3-gfp adg1{Delta} (KGY5093), spn3-GFP adg2{Delta} (KGY5094), and spn3-gfp adg3{Delta} (KGY5095) strains were grown at 32°C and visualized as in (Fig. 2D). (B) mid2{Delta} (KGY3135), eng1{Delta} (KGY5032), mid2{Delta} eng1{Delta} (KGY5467) and mid2{Delta} eng1{Delta} agn1{Delta} (KGY5505) cells were grown at 32°C and stained with DAPI to visualize DNA and Methyl Blue to visualize septa. (C) Quantification of the number of septa in cells from (B) in addition to wild-type (KGY246), agn1{Delta} (KGY5033), and eng1{Delta} agn1{Delta} (KGY5034) cells. In each case, 200 cells were examined. (D) spn3-gfp ace2{Delta} (KGY4774) containing the pREP41-mid2+ plasmid (pKG2208) was grown at 32°C for 18 hours in media lacking thiamine (–T). 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. Arrows indicate split ring structures that form after septation. Bars, 5 µm.