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. 2015 Mar 6;11(3):e1005036. doi: 10.1371/journal.pgen.1005036

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© 2015 Orgil et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 2 — A. Flowchart of the experimental design. B. Strain YIO81 with plasmid pRS406 (pGAL URA3), pIO88 (pGAL-SCC3 URA3) or pIO88R1 (pGAL-scc3-I358ins URA3) were grown in YEP lactic acid (2%) to mid-log phase and arrested in G1 using α-factor. Galactose was added to a final concentration of 2% for 1 h; cells were then released into the cell cycle and rearrested in G2/M by nocodazole. Sister chromatid cohesion was analyzed by using the GFP spot assay (n = 3). C. Strains analyzed in B were processed for chromatin spreads after cells were arrested in G2/M, as described in the materials and methods section. Cohesin was detected by indirect immunofluorescence with an anti-Mcd1 antibody (n = 2).