High through-put screen for <i>dRas</i>^<i>V12</i>-driven epithelial tumorigenesis in <i>Drosophila</i>.

(a) GFP+ eye disc cells in control, dRasV12, dRasV12;baz-RNAi (bazi), and dRasV12;dlg-RNAi (dlgi) flies. Ectopic GFP+ foci (arrowheads) in dRasV12;baz-RNAi (bazi), and dRasV12;dlg-RNAi (dlgi) observed on day 8 after egg laying (AEL) at 29°C. Magnifications X 8. (b) Normal and dRasV12-transformed eye discs at d5 AEL (top panels). Invasive disc tumors appearing upon co-expression of baz-RNAi or dlg-RNAi fuse with the brain lobes (arrows) in d8 AEL larvae (bottom panels). Magnifications X 25 (top panels) and X 40 (bottom panels). vnc, ventral nerve cord. (c) Quantification of larval arrest (% larval progeny at the indicated days AEL) and (d) independent test results for assay optimisation. The blue dotted line in (C) and the red line in (D) indicate the cut-off (Z-score 1.65) for phenotype detection. (e) Schematic of the screen design. Virgin driver line females (ey-Flp; Act5C>>GAL4, UAS-GFP/CyO, Tub-GAL80; UAS-dRasV12) were crossed to UAS-RNAi transgenic males. All crosses were performed at 29°C and parents removed after 4 days. Counts of the larval progeny were performed at two time points, d8 and d12 AEL, and compared to larval offspring from isogenic w1118 control males crossed to driver females.