10xstat92e gfp

The lines of jhamt^RNAi (103237 and 19172), Fpps^RNAi (104362), Sni^RNAi (106219 and 27342), Cyp305a1^RNAi (101644 and 51486), Met^RNAi (100638 and 10801), gce^RNAi (11176), tai^RNAi (15709), Jhe^RNAi (44049) and PTEN^RNAi (109278) were obtained from VDRC. Met^RNAi (61935), gce^RNAi (combination of 61852 and 26323 at the same time), tai^RNAi (36095), UAS-PI3K (8287), UAS-Rheb (9689), UAS-S6KII (8714), 10xSTAT92E-GFP (26197), UAS-EGFP-NiPp1 (23712) and Aug21Gal4 (30137) were obtained from Bloomington Stock Center. UAS Diap-1 was a gift from G. Morata. UAS-AstC was generated cloning the EcoRI-KpnI fragment from the cDNA clone RH36507 (DGC gold BDGP) into the pUAST vector. Transgene expression together with UAS-GFP was driven by esg-Gal4, Dl-Gal4, Su(H)-Gal4 or pros-Gal4. Gal4 activity was regulated by Tub > Gal80^ts. Flies were crossed at 17 °C, and two day old progeny was transferred to 29 °C for analysis. MARCM clones were generated by a 1hr heat shock at 37 °C of 2–5 days old females and were marked by the tubulin > Gal4 line driving the expression of UAS GFP (normal clones). Apc-Ras clones were generated as described previously^{15 (link)}.

Flies were raised on standard media. Crosses were raised at 25°C unless otherwise noted. 10xstat92E-GFP (BL#26197) [22 (link)], UAS-eGFP (BL#5430), UAS-eGFP (BL#5431), hsflp¹²²;;Ubi-RFP,FRT79 (made from BL#34498), y¹v¹hop^Tum/FM7c (BL#8492; referred to as hop^Tum-l), act5c-GAL4/CyO (BL#4414), UAS-MYR-RFP/CyO (BL#7118), hml-GAL4 (BL#30139) and stat⁰⁶³⁴⁶ (BL# 11681) were obtained from Bloomington Drosophila Stock Center, Bloomington, IN. UAS-hipk^RNAi (VDRC ID#108254, [23 (link)]) was obtained from Vienna Drosophila Resource Center, Vienna, Austria. Also used were dpp-GAL4/TM6B [24 (link)], os,y (a gift from Norbert Perrimon), UAS-Stat92E-GFP/Cyo and UAS-Stat92E-MYC/Cyo,wg-lacZ (a gift from James Castelli-Gair Hombria, [25 (link),26 (link)], PD-lacZ (a gift from Henry Sun; referred to as upd¹-lacZ hereon after, Tsai and Sun, 2004 [27 (link)]), ywhsflp,tub-GAL4,UAS-GFP,6X MYC-NLS; UAS-y+;tub-GAL80,FRT2A/TM6B (a gift from Gary Struhl), ywhsflp¹²²;sp/Cyo;TM2/TM6B, UAS-HA-hipk^1M, UAS-HA-hipk^3M, hipk⁴,FRT79/TM6B [28 (link)], UAS-HA-hipk^WT-attP40, UAS-MYR-HA-hipk-attP40, UAS-NLS-HA-hipk-attP40 (made in this study). act5c-GAL4/CyO and UAS-MYR-RFP/CyO were recombined to generate act5c-GAL4, UAS-MYR-RFP/CyO. hipk⁴, FRT79/TM6B and 10xstat92E-GFP/TM6B were recombined to generate hipk⁴, FRT79,10xstat92E-GFP/TM6B.

The Cg-gal4 (#7011), UAS-Dome-IR (#34618), UAS-mCD8-GFP (#5137), Canton-S (#64349) and 10XStat92E-GFP (#26197) stocks were obtained from Bloomington Drosophila Stock Center and maintained under standard conditions (25 °C, 50% relative humidity, 12/12-h light/dark cycles) in normal diet (ND, 0.15 M sucrose). For experiments, flies UAS-mCD8-GFP and UAS-Dome-IR were crossed to Cg-gal4 > UAS-mCD8-GFP, and embryos were collected on apple agar plates for 24 h. Early first instar larvae were transferred to an ND or high sucrose diet (HSD, 1 M sucrose) for 72 h and 120 h respectively at 29 °C.
The diets were prepared using Sigma reagents according to protocols previously described in the literature^{33 (link)}. For more details on the formulation of diets, review Supplementary Material Table 1.