Fig. 2. Disseminating Ras^V12-transformed cells undergo extensive remodeling of cell morphology.

a Representative cell morphologies from side and top view. Transgenes were induced for 1 day. Phalloidin signal visualizes VM (red). Scale bars, 10 µm. b Representative image of pHH3 (red) staining of Ras^V12 cells and quantification of phospho-histone H3 (pHH3) cells per midgut after 3 days of expression. Prior to staining, Ras^V12 was expressed for 2 days. Dotted line indicates the epithelial boundary. Scale bars, 10 µm. For quantification, day 1: N = 16 (esg^ts), N = 27 (esg^ts > Raf^gof), N = 6 (esg^ts > Ras^V12); day 2: N = 14 (esg^ts), N = 31 (esg^ts > Raf^gof), N = 8 (esg^ts > Ras^V12); day 3: N = 16 (esg^ts), N = 29 (esg^ts > Raf^gof), N = 6 (esg^ts > Ras^V12) biological replicates. Data are mean ± SEMs. c Delta (Dl) antibody staining. The Notch receptor ligand Dl—a marker of intestinal stem cells—was detected in Ras^V12 cells. Dotted line indicates the epithelial boundary. Ras^V12 cells are marked with GFP (green), and nuclei are stained with DAPI (blue). Scale bar, 10 µm. d Side view of Ras^V12 cell. Ras^V12 was induced for 2 days with esg^ts. Arrowheads point to protrusions formed across VM. Scale bar, 10 µm. e Side (xy) and orthogonal (yz and xz) views of Ras^V12 cell. Ras^V12 was expressed for 3 days. Scale bar, 10 µm.