Sugar and powdered yeast

Flies were reared at 25 °C in medium containing Nutri-Fly^TM Bloomington formulation (Genesee Scientific, San Diego, CA), Jazz mix (Fisher Scientific, Waltham, MA, USA), sugar and powdered yeast (Genesee Scientific) in an 8:5:1:1 ratio and made according to standard procedures. The following fly stocks were used in this study: UAS-Par-1, UAS-Par-1^T408A, and UAS-Par-1^RNAi (All gifts from Bingwei Lu, Stanford School of Medicine, Stanford, CA, USA^{25 (link)}), slmb^3A1 (Bloomington Stock Center)^{52 (link)}, olk^{1 (link)} and olk^{3 (link)} (a gift from Doris Kretzschmar, Oregon Health and Science University, Portland, OR, USA)^{53 (link)}. The following GAL4 lines were used: BG380-Gal4 (A gift from Aaron DiAntonio, Washington University Medical School, St. Louis, MO, USA)^{26 (link)}, and ELAV-GeneSwitch (Bloomington Stock Center)^{27 (link)}.

All fly lines were reared in medium containing Nutri-Fly^TM Bloomington formulation (Genesee Scientific, San Diego, CA), Jazz mix (Fisher Scientific, Waltham, MA, USA), sugar and powdered yeast (Genesee Scientific) in an 8:5:1:1 ratio and made according to standard procedures. To allow for the survival of homozygous trc¹ mutant larvae up to the early third-instar stage, flies had to be reared in bottles containing the medium described above at 18°C. We ensured that the bottles were optimally populated (~50–70 flies/bottle (max)). WT flies were either Canton S (CS) or CS outcrossed to w⁻, BG380.Gal4[35 (link)] or G7.Gal4[36 (link)] lines depending on the experiment. The following fly lines were obtained from the Bloomington Stock Center: trc¹[38 (link)], trc^Df: Df[3L]BSC445, wsp¹and UAS-Wasp[48 (link)], scar¹, gig¹⁰⁹[30 (link)] (Tsc2), Akt1⁰⁴²²⁶[69 (link)], and rictor^Δ42[70 (link)]. Expression of transgenic trc^RNAi lines: trc^GL01127P{TRiP.GL01127} [71 (link)] and trc^JF02961 P{TRiP.JF02961}[72 (link)] or p(UAS-trc-GFP)[41 (link)] was driven using the Gal4 lines mentioned above. BG380.Gal4 was obtained from Aaron DiAntonio, Washington University Medical School (St. Louis, MO, USA).