A) Selection of transgenic fly lines that express similar levels of ataxin-3 variants. Quantitative RT-PCR results from whole flies expressing the versions of UAS-ataxin-3 noted in the panel. At least 5 flies were used per genotype per experiment. Driver was sqh-Gal4 (ubiquitous expression31, 32). All flies were heterozygous for UAS-ataxin-3 and sqh-Gal4 transgenes. Red and blue arrows: lines that we chose for western blotting shown in panel B. Experiment performed independently in triplicate. Shown are mean ataxin-3 mRNA levels normalized to WT-1. Error bars: standard deviations. Flies were 1–3 days old.
B) Western blots from whole fly lysates expressing the indicated UAS-ataxin-3 constructs based on qRT-PCR data from panel A. Driver was sqh-Gal4. All flies were heterozygous for UAS-ataxin-3 and sqh-Gal4 transgenes, as in panel A. Ten or more flies per genotype were homogenized. Blots are representative of experiment performed independently in triplicate, with similar results. Flies were 1–3 days old.