Fig. 4

Regulation of dopaminergic levels of PARIS by PINK1 and parkin. a Immunoblot analysis and quantification of PARIS in flies expressing the indicated transgenes under the control of TH-Gal4 driver, N = 3. b Quantitative RT-PCR analysis in FACS sorted DA neurons showing PINK1 or parkin transcript levels upon TH-Gal4 mediated knockdown or overexpression of the respective genes. c Parkin functions downstream of PINK1 in a linear pathway to regulate PARIS levels in the DA neurons. Indicated transgenes expressed using TH-Gal4 driver, N = 4. d DA neuron loss in 30-day old flies expressing phosphodeficient PARIS double mutant (PARIS DM) is not rescued by dopaminergic overexpression of parkin or PINK1. N = 10 flies per indicated genotype. e Climbing defects in 30-day old TH > PARIS DM flies persist even under conditions of parkin or PINK1 overexpression in DA neurons. N = 60 flies per indicated genotype. f Dopaminergic overexpression of parkin or PINK1 does not attenuate accumulation of PARIS DM in DA neurons.TH-Gal4/+ flies served as control, N = 4. Quantitative data = mean ± SEM. One-way ANOVA, ns (non-significant), p > 0.05, *p < 0.05, **p < 0.01, ****p < 0.0001. Quantification of mRNA transcript levels relative to TH > GFP control from three independent FACS experiments each employing 50 fly brains of the indicated genotype. Data = mean ± SEM. Unpaired two-tailed Student’s t test *p < 0.05. See also Additional file 14, Figure S4