Fig. 2: The pathogenic Auxilin mutants develop age-dependent neurological abnormalities.
From: Parkinsonism mutations in DNAJC6 cause lipid defects and neurodegeneration that are rescued by Synj1
a Complementation table indicating viability/lethality of the indicated genotypes. L: lethal, SV: semi-viable and V: viable. b–c. Images of Western blots from brain lysates prepared from 5, 15 and 30DO flies of indicated genotypes labelled with anti-DNAJC6/Auxilin and anti-GAPDH (loading control) and quantification of Auxilin protein levels. Values are relative to GAPDH and are expressed as a fraction of protein levels in dAuxWT/WT and dAuxWT/F. Bars show the mean ± SEM, points show individual values and n≥3. Two-way ANOVA, with Bonferroni’s post hoc test. *p < 0.05, **p < 0.01 and ***p < 0.001. d–d’. Quantification of the negative geotaxis assay of 5DO, 15DO or 30DO flies of the indicated genotypes (d) and representative images of 15DO flies after 30 s (d’). Graphs represent the fraction of flies crossing the 4 cm mark. Note that most dAuxWT/F956X and all dAuxRG/F956X flies die within 25-days-post-eclosion (red crosses). Bars show mean ± SEM. Points represent the mean of three trials each. One-way ANOVA, Dunnett’s multiple comparison test. ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. †dead at this age. e–e’. Quantification of seizures following vortexing in 5DO flies of indicated genotypes (e) and representative images of flies after vortex-stimulation (e’). Graphs represent the fraction of flies that were unable to stand 10 s after stimulation. Bars show mean ± SEM. Points represent single trials with 10 flies each. Red arrows: flies that are not able to stand. One-way ANOVA, Dunnett’s multiple comparison test. ns: not significant, ****p < 0.0001.
