Fly lines w1118, FRT82, LabialRNAi (BL26753), UAS-mcherry (BL38245), UAS-TkvQD (BL36536), UAS-Dpp (BL1486), UAS-Ubx (BL911), Ubx1 (BL529), UbxRNAi line 1 (BL31913), UbxRNAi line 2 (BL34993), DadRNAi (BL33759) were obtained from Bloomington Drosophila Stock Center. esg-Gal4, UAS-GFP was a gift from Shigeo Hayashi; FRT82B, Dad212 from Hannele Ruohola-Baker; Btl-Gal4ts, UAS-GFP from Dirk Bohmann; Dad::nlsGFP from Georgios Pyrowolakis; Su(H)-GBE-lacZ from Sarah Bray; esgtsF/O (esgGal4, tubG80ts, UAS-GFP; UAS-flp, act > STOP > Gal4) from Huaqi Jiang; UAS::Dad from Thomas Kornberg; NP1::Gal4 from Dominique Ferrandon; MARCM82 (hsFlp; tub-Gal4, UAS-GFP; FRT82, tubGal80) from Norbert Perrimon.
Flies were cultured on yeast/molasses-based standard fly food (Recipe: 10 L H2O, 138 g agar, 220 g molasses, 750 g malt extract, 180 dry yeast, 800 g corn flour, 100 g soy flour, 62.5 ml propionic acid, 20 g Methyl 4-Hydroxybenzoate, and 72 ml ethanol) at 25 °C with a 12 h light/dark cycle. For TARGET (tubGal80ts) experiments, flies were raised at 18 °C to allow Gal80 to inhibit Gal4, and 3–4 days after eclosion shifted to 29 °C to inhibit Gal80 and to allow Gal4 to drive UAS-linked transgene expression.
Flies were cultured on yeast/molasses-based standard fly food (Recipe: 10 L H2O, 138 g agar, 220 g molasses, 750 g malt extract, 180 dry yeast, 800 g corn flour, 100 g soy flour, 62.5 ml propionic acid, 20 g Methyl 4-Hydroxybenzoate, and 72 ml ethanol) at 25 °C with a 12 h light/dark cycle. For TARGET (tubGal80ts) experiments, flies were raised at 18 °C to allow Gal80 to inhibit Gal4, and 3–4 days after eclosion shifted to 29 °C to inhibit Gal80 and to allow Gal4 to drive UAS-linked transgene expression.