Figure 4 | Cell Death & Differentiation

Figure 4

From: The Ca2+/Mn2+ ion-pump PMR1 links elevation of cytosolic Ca2+ levels to α-synuclein toxicity in Parkinson’s disease models

Figure 4

Expression of αSyn causes a slight upregulation of PMR1 and CCH1 mRNA levels. (a) Representative micrographs of yeast cells expressing endogenously GFP-tagged Pmr1p in combination with αSyn or corresponding vector control at indicated time points after induction of αSyn expression. (b) Western blot analysis of cells described in (a) at indicated time points after induction of αSyn expression. Blots were probed with antibodies against GFP to detect Pmr1p-GFP fusion protein, against FLAG-epitope to detect FLAG-tagged αSyn and against glyceraldehyd-3-phosphate dehydrogenase (GAPDH) as loading control. (c and d) Q-PCR-based quantification of PMR1 mRNA levels (c) and of CCH1 and MID1 mRNA levels (d) in WT cells expressing αSyn or harbouring the empty vector control for 14 h or 24 h, respectively, normalized to actin mRNA levels. Means±S.E.M., n=6–9; *P<0.05. (e) Quantification of ROS accumulation (DHE→Eth) in WT yeast cells and indicated deletion mutants expressing αSyn for 16 h or 24 h using a fluorescence reader. Mean±S.E.M., n=6; ***P<0.001 and *P<0.05

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