Figure 6

Expression of Pmr1p restores αSyn cytotoxicity and supresses manganese toxicity of PMR1-deficient cells. (a) Spotting assays of WT and Δpmr1 yeast cells expressing αSyn or harbouring the vector control. Cells were grown for 24 h in galactose media and spotted in fivefold serial dilutions onto glucose (αSyn expression repressed) and galactose (αSyn expression induced) agar plates supplemented or not with 2 mM or 4 mM Mn²⁺, respectively. (b) Spotting assays of WT and Δpmr1 yeast cells expressing either αSyn or Pmr1p alone or in combination or harbouring the corresponding vector controls. Cells were grown for 24 h in galactose media and spotted in fivefold serial dilutions onto glucose (Pmr1p and αSyn expression repressed) and galactose (Pmr1p and αSyn expression induced) agar plates supplemented or not with 4 mM Mn²⁺. (c) Quantification of clonogenic survival of cells described in (b) after plating on galactose agar plates supplemented or not with 4 mM Mn²⁺. Both Pmr1p as well as αSyn expression are driven by a galactose promoter. Mean±S.E.M., n=8–12; ***P<0.001 and **P<0.01; NS, not significant. Unless otherwise specified, asterisks indicate significances to similarly treated, isogenic control cells harbouring both empty vectors