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. 2013 Jun 15;24(12):1904–1917. doi: 10.1091/mbc.E12-10-0775

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© 2013 Tebbs and Pollard. 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 1: — Rng2-D5 cells assemble contractile rings more slowly at a semipermissive temperature than do wild-type cells. (A) Schematic of Rng2p domains. (B) Western blots with antibodies to GFP or cofilin of lysates of cells expressing mEGFP-Rng2p or mEGFP-Rng2-D5 at the permissive (25°C), semipermissive (32°C), or restrictive (36°C) temperature. (C) Time series of fluorescence micrographs at 2-min intervals at 32°C of wild-type (top row) and rng2-D5 (bottom row) cells expressing Sad1-CFP (red) to mark spindle pole bodies and YFP-Cdc15p (green) to mark nodes and contractile rings. Time in minutes is shown in white, with spindle pole body separation defined as time zero. (D) Average times (±1 SD) required for wild-type (open bars) and rng2-D5 (filled bars) cells to assemble (n = 27 and 9, respectively), mature (n = 25 and 21, respectively), and constrict (n = 21 and 19, respectively) contractile rings using YFP-Cdc15p as the marker for contractile rings. (E) Time series of fluorescence micrographs at 2-min intervals at 32°C of cells expressing Sad1-RFP (red) and (top) either mEGFP-Rng2p or (bottom) mEGFP-Rng2-D5 (green). Scale bars, 5 μm.