Fig. 2

RIZ2 overexpression Increases DLD1 3D-organoid growth and cell migration. A Representative images of stable transfected DLD1 cells in miniaturized 3D cultures in extracellular matrix (three independent experiments, each performed in duplicate). Scale bar, 100µ. Graphs illustrate the relative organoid size (area), which was calculated using the Application Suite Software and expressed as a relative fold increase at 8, 14 and 20 days over the organoid area calculated at 4th day. *p < 0.05 vs. control cells. B DLD1-pEGFP cells and DLD1-pEGFP_hRIZ2 were wounded and left for 18 h. Phase-contrast images are representative of three different experiments, each in duplicate. C The wound area was calculated using Leica Suite Software. Data are represented as % of residual wound width. The standard deviations were < 0.05 for each experimental condition (left graph). In middle and right graphs, DLD1-pEGFP cells and DLD1-pEGFP_hRIZ2 were allowed to migrate (middle) or invade (right) for 7 h or 24 h, respectively. They were scored as described in “Methods” section and data were expressed as fold increase. Means and SDs are shown. n = 3. *p < 0.05 for the indicated experimental points versus the corresponding 0 time cells