(A) Confocal images of third instar wing imaginal discs expressing ptc > GFP plus indicated transgenes (green) and rhodamine-phalloidin (purple) fluorescence in XY maximum projection images and (B) XZ single slices below (posterior to the right). (C) Quantification of ptc > GFP stripe width (in µm; mean ± SEM: ptc > GFP [26.4 ± 1.3, n = 6], ptc > GFP, Dnhe2 [27.4 ± 4.0, n = 6], ptc > GFP, RasV12 [43.8 ± 4.5, n = 6], ptc > GFP, RasV12, Dnhe2 [60.7 ± 3.9, n = 7]). (D) Quantification of total area of wing imaginal discs (in 103 pixels; mean ± SEM, ptc > GFP [137.3 ± 9.6, n = 9], ptc > GFP, Dnhe2 [107.8 ± 8.9, n = 9], ptc > GFP, RasV12 [93.5 ± 4.1, n = 12], ptc > GFP, RasV12, Dnhe2 [196.5 ± 41.4, n = 8]). (E) Invasive cells were identified as GFP-positive cells that were isolated from other GFP-positive cells in three-dimensional tissue architecture. For each example, shown are XY (upper left), YZ (upper right) and XZ (lower left) single confocal slices. Invasive phenotypes were rarely seen in ptc > GFP, ptc > GFP, Dnhe2 discs, or ptc > GFP, RasV12 wing discs. In ptc > GFP, RasV12, Dnhe2 discs, examples of basal stripe expansion (4/7), multiple single, invasive cells (5/7 discs) and streams of invading cells (2/7) were seen. (*p < 0.05, **p < 0.01, ***p < 0.001).