All fly crosses were performed at 25°C in standard laboratory conditions. For the HRP–Boss screen, w; FRT82neo males treated with 25 mM ethyl methanesulfonate (M0880; Sigma-Aldrich) were mass mated with w/w; TM3, Sb/TM6B, Hu, Tb virgins. Individual progeny were backcrossed to w; TM3, Sb/TM6B, Hu, Tb flies to establish lines. Several w; FRT82neo, mutant/TM6B, Hu, Tb males from each line were then mated to ey-FLP; P{w+, HRP-Boss}; FRT82neo, arm-lacZ females. Eye discs were dissected from Tb+ third instar larval progeny and stained for HRP and β-galactosidase activities.
Mitotic clones in adult retina were generated using hs-FLP1; FRTneo82B, P{w+}96A (Xu and Rubin, 1993 (link)). Mitotic clones in larval eye discs were generated using ey-FLP; FRT82neo, GMR-myrGFP-3R (see below). To facilitate exogenous protein expression in larval Garland cells, UAS-derived transgenes (UAS-GFP–rab7 and UAS-GFP–Clc) were driven with Act5C-GAL4 lines.
UAS-myc::Hsc70-C1 and alleles of Hsc70-4 were obtained from Spyros Artavanis-Tsakonas (Massachusetts General Hospital/Harvard Medical School, Boston, MA). HRP-Boss flies were obtained from Helmut Kramer (University of Texas Southwestern, Dallas, TX). UAS-GFP–rab7 flies were obtained from Marcos A. González-Gaitán (Max-Planck Institute, Dresden, Germany). Act5C-GAL4 (No. 4414), Sev-GAL4 (No. 5793), and sca-GAL4109–68 (No. 6479) were obtained from the Bloomington Drosophila stock center (Bloomington, IN).
Mitotic clones in adult retina were generated using hs-FLP1; FRTneo82B, P{w+}96A (Xu and Rubin, 1993 (link)). Mitotic clones in larval eye discs were generated using ey-FLP; FRT82neo, GMR-myrGFP-3R (see below). To facilitate exogenous protein expression in larval Garland cells, UAS-derived transgenes (UAS-GFP–rab7 and UAS-GFP–Clc) were driven with Act5C-GAL4 lines.
UAS-myc::Hsc70-C1 and alleles of Hsc70-4 were obtained from Spyros Artavanis-Tsakonas (Massachusetts General Hospital/Harvard Medical School, Boston, MA). HRP-Boss flies were obtained from Helmut Kramer (University of Texas Southwestern, Dallas, TX). UAS-GFP–rab7 flies were obtained from Marcos A. González-Gaitán (Max-Planck Institute, Dresden, Germany). Act5C-GAL4 (No. 4414), Sev-GAL4 (No. 5793), and sca-GAL4109–68 (No. 6479) were obtained from the Bloomington Drosophila stock center (Bloomington, IN).
