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. 2016 Jun 2;44(16):7722–7741. doi: 10.1093/nar/gkw506

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 8. — The ATPase activity of Pch2 is required for its checkpoint function. (A) Schematic representation of the Pch2 protein indicating the AAA+ domain, the conserved Walker A and Walker B motifs, and the mutations introduced at both sites. (B) Western blot analysis of Pch2, Pch2-K320A or Pch2-E399Q production (detected with anti-HA antibodies) during meiosis. (C) Dityrosine fluorescence assay. (D) Time course of meiotic nuclear divisions; the percentage of cells containing two or more nuclei is represented. (E) Western blot analysis of Hop1^T318 phosphorylation and Mek1 activation. PGK was used as a loading control. Strains are: DP1151 (wild type), DP1164 (pch2Δ), DP1163 (pch2-K320A), DP1287 (pch2-E399Q), DP1152 (zip1), DP1161 (zip1 pch2Δ), DP1162 (zip1 pch2-K320A) and DP1288 (zip1 pch2-E399Q).