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. 2011 Jun 15;22(12):1985–1996. doi: 10.1091/mbc.E10-06-0545

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© 2011 Lin 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 2: — Scc2 is required for nuclear division and sister-chromatid cohesion during meiosis. (A) Depletion of Scc2 during yeast meiosis. Wild-type (3050) and P_CLB2-SCC2 (HY1336) cells were induced to undergo synchronous meiosis, aliquots were withdrawn at indicated time points, and samples were prepared for immunoblot. Scc2-3HA was detected by an anti-HA antibody (16B12); Dmc1 was detected by a Dmc1-specific polyclonal antibody. Dmc1 is a meiosis-specific protein, indicating meiosis induction. The level of β-tubulin served as a loading control. (B) Nuclear division. Yeast cells were induced to undergo synchronous meiosis, and aliquots were withdrawn at indicated time points and fixed in 1% formaldehyde for 1 h. Cells were stained with DAPI; nuclear division was determined by fluorescence microscopy. WT, NH144; P_CLB2-SCC2, HY1336; spo11-Y135F, HY1499; P_CLB2-SCC2 spo11-Y135F, HY1975. (C) Sister-chromatid cohesion assay. A tandem array of tetO was inserted at the URA3 locus of one homologue of chromosome V, and the separation of GFP dots was determined by fluorescence microscopy. WT, HY1294C; P_CLB2-SCC2, HY2113; ndt80Δ, HY2130; P_CLB2-SCC2 ndt80Δ, HY2115.