Figure 4

(a–c) eye-FLP-MARCM experiments showing GFP⁺ tissues (close-up view in bottom panels a′, b′, c′) with the genotypes dNek2GFP (a, a′), Csk−/−; Ras^V12(b, b′) and dNek2GFP; Csk−/−; Ras^V12 (c, c′). dNek2GFP; Csk−/−; Ras^V12 larvae show a significant increase in primary tumor mass (arrows in c and c′) as well as appearance of secondary tumors (asterisks in c) at a distance from the primary tumors. (d–k) eye-FLP-MARCM experiments showing that dNek2GFP; Csk−/−; Ras^V12 tissues upregulate Diap1 (e, e′), Mmp1 (g, g′), Wg (i, i′) and CycE (k, k′), in comparison with Csk−/−; Ras^V12 tissues (d, d′, f, f′, h, h′, j, j′). Epifluorescence′ images were taken at equivalent settings for each channel. (l) Tumor cell injection experiments. GFP⁺ eye-FLP-MARCM tissues obtained from third instar larvae were dissociated and injected into the dorsal notum of w- adult flies. Injection of dNek2GFP cells (n=200) or Csk−/−; Ras^V12 cells (data not shown) did not lead to GFP⁺ cells seeding into the host fly. But injection of dNek2GFP; Csk−/−; Ras^V12 cells led to appearance of distinct GFP⁺ foci within 10 days of injection (20 out of 180 injected flies showed seeding injected tumor cells). Site of injection is visible as a dark scar (arrow) and three examples are shown with tumor foci in parts of the body away from the injection site (asterisks).