Fig. 1
Analysis and quantification of the effects induced by expression of SCA3polyQ78. a Representative pictures of a control eye, and eyes expressing SCA3polyQ27 or SCA3polyQ78. Expression of SCA3polyQ78 but not SCA3polyQ27 in Drosophila eyes resulted in loss of pigment (“depigmented”), as well as the presence of necrotic patches (“necrotic”, indicated with arrow). b Quantification of the fraction of the eyes as shown in a that have a normal appearance, display a loss of pigment or contain necrotic patches; the fraction of SCA3polyQ78 eyes containing necrotic patches was used as a quantitative measure for degeneration. Data are representative of at least three independent experiments. SEM = 0.863. c Western blot analysis of fly head lysates expressing eye-specific, HA-tagged SCA3polyQ27, or SCA3polyQ78. A fraction of SCA3polyQ78 but not SCA3polyQ27 was SDS-insoluble. Tubulin was used as a control for equal loading. d Expression of mCD8-GFP in the eye to analyze SCA3polyQ78-induced eye degeneration and analysis of SCA3polyQ78 expression on the localization of astrocytes (expressing RFP). Top panel: GFP fluorescence of a representative control eye or eyes coexpressing SCA3polyQ27 or SCA3polyQ78. Bottom panel: the localization of myr-RFP-expressing (RFP) astrocytes was analyzed in control eyes or eyes expressing SCA3polyQ27, or SCA3polyQ78. e Lysates of control flies or flies expressing mCD8-GFP in the eyes and myr-RFP in astrocytes together in the absence or presence of SCA3polyQ27 or SCA3polyQ78 were analyzed for expression of HA-tagged SCA3, GFP, and RFP. Tubulin was used as an equal loading control. Genotypes in a–c: control, GMR-QF2/+. SCA3polyQ27, GMR-QF2/+; QUAS-SCA3polyQ27/+. SCA3polyQ78, GMR-QF2/+; QUAS-SCA3polyQ78/+. Genotypes in d, top panel: control, GMR-QF2/+; QUAS-mCD8-GFP/+. SCA3polyQ27, GMR-QF2/+; QUAS-SCA3polyQ27/+; QUAS-mCD8-GFP/+. SCA3polyQ78: GMR-QF2/+; QUAS-SCA3polyQ78/+; QUAS-mCD8-GFP/+. Bottom: control, GMR-QF2/+; alrm-Gal4::UAS-myr-RFP/+. SCA3polyQ27, GMR-QF2/+; alrm-Gal4::UAS-myr-RFP/QUAS-SCA3polyQ27. SCA3polyQ78: GMR-QF2/+; alrm-Gal4::UAS-myr-RFP/QUAS-SCA3polyQ78. Genotypes in e: as in d except that, where indicated, both QUAS-mCD8-GFP (eyes) or alrm-Gal4::UAS-myr-RFP (astrocytes) were expressed