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. Author manuscript; available in PMC: 2015 Feb 21.
Published in final edited form as: Nat Commun. 2014 Aug 21;5:4638. doi: 10.1038/ncomms5638

Figure 5. UbS2 mutation leads to reduced ataxin-3 protein levels in Drosophila.

Figure 5

A) Quantitative RT-PCR results from whole flies expressing the noted versions of UAS-ataxin-3 driven by sqh-Gal4. All flies were heterozygous for UAS-ataxin-3 and sqh-Gal4 transgenes. Red arrows: WT and UbS2* lines that have comparable ataxin-3 mRNA levels. Blue arrows: UbS2* lines that have markedly higher ataxin-3 mRNA levels than WT versions. Experiment performed independently in triplicate, utilizing at least 5 flies per genotype per experiment. Shown are mean ataxin-3 mRNA levels normalized to WT-1. Error bars: standard deviations.

B) Left: Western blots from whole flies based on qRT-PCR results from panel A. At least 5 flies were homogenized per genotype. Driver was sqh-Gal4. All flies were heterozygous for UAS-ataxin-3 and sqh-Gal4 transgenes, as in panel A. Note that for this blot 1.5× more lysate was loaded for the line that expresses UbS2*-2 to enable visualization of ataxin-3 protein in this line by western blotting without saturating the signal from other lysates. Right: Means of ataxin-3 signal quantified from blots on the left and other independent experiments. P values are from Student T-test (WT-3 and UbS2*-2) and ANOVA/Tukey (the other lines). Error bars: standard deviations. N=3 independently conducted experiments. Flies were 1–3 days old.