Fig. 2. Dosage dependence of the phenotype. (A,B) Flies carrying a UAS-eyHD transgene in addition to the dpp^blink-Gal4 transgene. A and B are two independent transgenic lines of the same transgene showing ectopic eyes (arrowheads) obtained to a different degree of strength. (C) Western blot analysis using a rabbit -EY antibody on third instar leg discs expressing the transgenes used for the phenotypes obtained in panels A and B. The strength of the phenotype correlates with the amount of ectopically induced protein. Asterisk marks the protein expressed at lower level corresponding to the ectopic eyes in panel A. Molecular weight marker is indicated at the right. Lanes 1 and 2, yw control wing and leg discs, respectively; lanes 3-5, leg discs expressing the various ey constructs by dpp^blink-Gal4. (Lane 3) Misexpression of eyHD, transgenic line of panel A. (Lane 4) Misexpression of eyHD, transgenic line of panel B. (Lane 5) Misexpression of full-length ey. (D-F) Temperature dependence of the Gal4 system. (D,E) Flies carrying the UAS-toy transgene and the dpp^blink-Gal4 transgene in an ey² mutant background raised either at 18°C (D) or at 25°C (E). Ectopic eyes were never observed on flies raised at 18°C, in contrast to flies raised at 25°C. (F) Western blot analysis with a rabbit -quail-PD antibody of third instar leg discs expressing the various Pax6 transgenes under the control of the dpp^blink-Gal4 transgene in a wild-type background. Pax6 proteins are marked by asterisks. Molecular weight marker is indicated at the right. (Lanes 6-11) Misexpression of ey (lane 6), eyHD (lane 7), toy (lane 8), toyHD (lane 9), seyHD (lane 10) and seyHD (lane 11).