Fig. 5

ZD1839 greatly neutralized RIZ2 oncogenic properties. In A–D, DLD1-pEGFP and DLD1-pEGFP_hRIZ2 stable cell lines were used. Cells were untreated or treated with ZD1839 (ZD) and analyzed for cell proliferation (A) through WST-1. B Cell lines were used in miniaturized 3D cultures in extracellular matrix (ECM), as reported in Methods. Four days after cells embedding in Matrigel (starting) representative images were acquired as described in Methods and cells were unchallenged or challenged with ZD1839 (ZD) for 18 days. Shown are phase-contrast images captured on the 18th day. Scale bar, 100 µ. Graphs on the right show the analysis of organoid size from cell lines unchallenged or challenged with ZD. The size of organoids was calculated using Leica suite software. For each cell line, 3 different experiments were done. Means and SDs are shown. Cell migration was analyzed through wound scratch assay (C) or collagen-coated transwells (D, left panel) and cell invasiveness trough Matrigel-coated transwells (D, right panel) as described in Figs. 1, 2 legends and Methods section. In A–D means and SDs are shown. *p < 0.05; **p < 0.001; ***p < 0.0005; ****p < 0.0001; ns not significant