TRAP1 Mitigates a-Synuclein Toxicity Figure 2. TRAP1 overexpression mitigates detrimental effects induced by neuronal [A53T]a-Synuclein expression. (A) Overexpression of [A53T]a-Synuclein under control of ddc-GAL4 resulted in reduction of DA in fly heads at 4 weeks, which was potentiated by TRAP1 deficiency (TRAP1[KG]/+;ddc.A53T/+), but mitigated by TRAP1 overexpression (ddc.A53T/hTRAP1). (B) ddc.A53T flies display a reduction of TH-positive neurons, which was potentiated by TRAP1 deficiency, but rescued to control levels by TRAP1 overexpression. (C) In negative geotaxis assays ddc.A53T flies displayed a time-dependent decline in locomotion. Reduction of TRAP1 enhanced the inability to climb (although not significant), while overexpression of hTRAP1 provided a significant rescue effect (comparison of ddc.A53T/+ vs ddc.A53T/hTRAP1 at 4 weeks: p,0.05). Statistics in (A, B): ANOVA followed by Newman-Keuls Multiple Comparison Test; (C): 2-way ANOVA followed by Bonferroni post-hoc tests. Displayed are biologically relevant comparisons. *p,0.05; **p,0.01; ***p,0.001; ns = not significant. (D) Alterations in TRAP1 levels did not influence PolyQ- induced rough eye phenotypes. Light micrographs of external eye structures show that PolyQ-induced REP was suppressed by parallel expression of HSP70. In contrast, neither overexpression of hTRAP1 or silencing of endogenous TRAP1 by RNAi had an obvious impact on external eye structure. Expression of mitochondrial localized GFP (mito-GFP) served as control. doi:10.1371/journal.pgen.1002488.g002 PLoS Genetics | www.plosgenetics.org 4 February 2012 | Volume 8 | Issue 2 | e1002488