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

Figure 3

PA and PI(3,5)P2 mediated protection to mitochondrial stress occurs via ATP13A2. SHSY5Y cells stably expressing FLUC, shRNA for KD of ATP13A2, ATP13A2, and full length mutants of ATP13A2 with mutated putative lipid binding sites (LBS1–3) were exposed to rotenone (Rot, 1 μM) in the presence or absence of 1 h pretreatment with the pharmacological inhibitors of PIKfyve lipid kinase (((a) and (b)) YM-201636, 200 nM) or phospholipase D (((c) and (d)) FIPI, 100 nM). Following 24 h exposure to rotenone, cells were assessed for cell viability ((a) and (c)) or death ((b) and (d)) by, respectively, MUH and propidium iodide (PI) assays. Data are the mean of 3 independent experiments ± SD. statistical differences between FLUC and WT-OE/sh-ATP13A2/LBS1/LBS2/LBS3/LBS1.2.3, statistical differences within cell line following treatment with inhibitor (1 mark, ; 2 marks, ; 3 marks, ) (ANOVA with Bonferroni post hoc test).

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