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. 2009 Feb 1;20(3):1030–1047. doi: 10.1091/mbc.E08-06-0637

Figure 5.

Figure 5.

rec8-6A and rec8-29A cells form sister chromatid cohesion. (A and B) Wild-type (A21230, ■), rec8-17A (A21234, ▴), and rec8-29A (21232, ·), all carrying GAL-NDT80 and GAL4-ER fusions, were induced to sporulate in the absence of βE. βE was added after 6 h, and samples were taken to determine the percentage of cells with unassembled meiotic spindles (A) and of cells with metaphase I spindles (B). n = 200 cells counted per time point per strain. (C and D) Wild-type (A1972, ■), rec8Δ (A3528, ▴), rec8-29A (A14385, ·), and rec8-6A (A15042, □) cells were induced to sporulate. At the indicated times, samples were taken to determine the percentage of cells with unassembled spindles (A) and of cells with metaphase I spindles (B). n = 200 cells counted per strain per time point. (E) spo11Δ spo13Δ (A21466, A21467, and A21468), spo11Δ spo13Δ rec8Δ (A21472, A21473, and A21474), spo11Δ spo13Δ rec8-29A (A21469, A21470, and A21471), and spo11Δ spo13Δ rec8-6A (A21463, A21464, and A21465) cells carrying a tet repressor-GFP fusion construct and one heterozygous tandem tet operator array inserted at the LEU2 locus were induced to sporulate on plates. After 24 h of sporulation, dyads were scored for sister chromatid segregation by fluorescence microscopy. Cells in which sister chromatids properly segregated apart are represented by the white portion of the bar, whereas cells in which sister chromatids segregate together are represented by the gray portion of the bar. n = 100 cells counted per strain in three independent isogenic diploid strains. Error bars, SD. (F) Wild-type cells with homologous LYS2 dots (A9828, ■), rec8-6A cells with homologous LYS2 dots (A21669, ▴, rec8-29A cells with homologous LYS2 dots (A16412, ·), and wild-type cells with nonhomologous LEU2/LYS2 dots (A11474, □), all deleted for NDT80, assayed for pairing as described in Figure 1A. Note that the same nonhomologous dot control strain is shown in F and G, because the experiments were performed in parallel. (G) Wild-type cells with homologous LEU2 dots (A5111, ■), rec8-6A cells with homologous LEU2 dots (A21670, ▴), rec8-29A cells with homologous LEU2 dots (A21668, ·), and wild-type cells with nonhomologous LEU2/LYS2 dots (A11474, □), all deleted for NDT80, were assayed for pairing as described in Figure 1A.