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. 2012 Apr 1;23(7):1181–1195. doi: 10.1091/mbc.E11-09-0800

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© 2012 Ye et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — Overexpression of Mid1 rescues gef2∆ plo1-ts18 defects in division-site positioning. (A) Overexpression of Mid1 under different promoters partially rescues septum-positioning defect in gef2∆ plo1-ts18 cells. gef2∆ plo1-ts18 (JW3078), 2x mid1 gef2∆ plo1-ts18 (an extra copy Pmid1-mid1 integrated at the leu1 locus in addition to the native mid1; JW3242), and gef2∆ plo1-ts18 Purg1-mid1 (integrated at the mid1 locus and induced in media with uracil; JW3237) were grown in YE5S medium at 25°C for 24 h. Top, DIC; bottom, calcofluor staining. Septum positioning in calcofluor-stained cells was quantified (right). Mean ± SD from three independent experiments. (B) Two copies of Mid1 partially rescue Mid1 ring formation in gef2∆ plo1-ts18 cells expressing Mid1-mECitrine and CFP-Atb2. Strains JW3281 and JW3383 grown at 25°C. The percentage of mitotic cells (with a spindle) with a Mid1 ring was quantified from two experiments. (C) Mid1-Cdr2 fusion protein partially rescues septum-positioning defect in gef2∆ plo1-ts18 cells. Left, DIC; middle, calcofluor staining; right, Mid1-mEGFP (top, JW3560) or Mid1-Cdr2-mEGFP (bottom, JW3426). Septum positioning in calcofluor-stained cells was quantified from two experiments. Bars, 10 μm.