Protection against Mitochondrial and Metal Toxicity Depends on Functional Lipid Binding Sites in ATP13A2

Figure 2

LBS mutations prevent ATP13A2-mediated cytoprotection. (a) ATP13A2’s protective response at 24 h exposure to rotenone (Rot, 1 μM) and the effect of pharmacological inhibition of PIKfyve lipid kinase with YM-201636 (PIK or P, 200 nM) and inhibition of PLD with FIPI (F, 100 nM) on cell death were assessed via a propidium iodide (PI) based assay. statistical differences between FLUC and WT-OE/sh-ATP13A2, statistical differences within cell line following treatment with inhibitor (1 mark, ; 2 marks, ; 3 marks, ) (ANOVA with Bonferroni post hoc test). ((b)–(e)) Stable cell lines were exposed to 1 μM rotenone ((b)-(c)) or 50 μM MPP+ ((d)-(e)) for 24 h and cell death was assessed by PI stained flow cytometry, whereas cell viability was assayed by the MUH protocol. Data are the mean of 3 independent experiments ± SD. statistical differences between FLUC and WT-OE/sh-ATP13A2/LBS1/LBS2/LBS3/LBS1.2.3 (1 mark, ; 2 marks, ; 3 marks, ) (ANOVA with Bonferroni post hoc test).

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