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. 2011 Jan 18;28(7):2005–2017. doi: 10.1093/molbev/msr019

FIG. 7.

FIG. 7.

Mdj2 does not compensate for lack of the IMS domain of Pam18 in the absence of oxygen. (A) Cells lacking Mdj2 have no anaerobic growth defect. Ten-fold serial dilutions of wild-type (WT) and mdj2-Δ cells were spotted on SC + Erg/TW plates and grown in the presence (+O2) or absence (−O2) of oxygen at the indicated temperatures. (B) No synthetic genetic interaction between mdj2-Δ and pam18Δ1–60 mutations. The indicated combinations of mutants were grown in the presence (+) or absence (−) of oxygen as described in A. In the case of PAM18, the chromosomal copy was deleted, and the control WT Pam18 (WT) and Pam18Δ1−60 (Δ1−60) were expressed from the pRS315 plasmid. (C) Overexpression of Mdj2 does not rescue the anaerobic growth defect of pam18Δ1−60 cells. (Left) pam18-Δ cells expressing either full-length Pam18 (WT) or Pam18Δ1−60 (Δ1−60) from pRS315 as well as harboring either p416 having MDJ2 under the control of the GPD promoter (MDJ2↑) or p416 control vector (empty) were grown in the presence (+O2) or absence (−O2) of oxygen. SC-Ura-Leu dropout media with Ergosterol and Tween 80 supplement (SC-Ura-Leu + Erg/Tw) were used to ensure plasmid retention. Cells were grown at 34 °C aerobically for 2 days or anaerobically for 2.5 days. (Right) Whole cell lysates prepared from cells cultured on SC-Ura-Leu + Erg/Tw at 30 °C under anaerobic condition were used for the immunoblot analysis with antibodies against Mdj2 and Tim23, as the loading control.