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. 2016 Sep 22;45(1):106–114. doi: 10.1093/nar/gkw818

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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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Oxs1-mediated diamide tolerance requires Pap1. (A) Cells of 10-fold serially diluted containing empty vector pART1 (EV) or pART1 expressing oxs1⁺ spotted on EMM selective media without or with diamide or Cd. (B) GST-Pap1 or GST bound to GST affinity resin incubated with His-Oxs1. Pull-down fractions analyzed by western blotting with anti-His antibody. (C) GST-Oxs1 or GST bound to GST affinity resin incubated with His-Pap1. Pull-down fractions detected with anti-His antibody. (D) Protein extracts from strain producing HA-Pap1, FLAG-Oxs1 or both were treated without or with diamide (1 mM, 3 h), Cd (0.1 mM, 3 h) or H₂O₂ (0.2 mM, 5 min), immunoprecipitated with anti-HA antibody and detected by anti-HA or anti-FLAG antibody. Input fractions represent 2% of His-tagged proteins in pull-down assays. Numbers left of panels B, C and D indicate size markers in kD.