Fig. 1
ID8 and ID8-KRAS cells in 2D and 3D conditions in vitro. ID8 and ID8-KRAS cells (1 × 106) were incubated for 48 h in 2D or 3D culture. a Representative microscopic images of cancer cells in the 2D and 3D culture conditions at 100 × magnification. b-c Number of viable cells for ID8 and ID8-KRAS in the 2D (b) and 3D conditions (c). Cultured cells were collected, and live, trypan blue-excluding cells were counted to determine the number of viable cells. Experiments were repeated at least thrice. The values shown represent the mean ± SEM (* p < 0.05). d-e ID8-GFP and ID8-KRAS-GFP cells (1 × 106) were incubated for 48 h in 2D or 3D culture. For 5-ethynyl-2′-deoxyuridine (EdU) proliferation analysis, cells were exposed to 10 μM EdU for 2 h before fixation. EdU was labeled with Alexa Fluor 647. EdU uptake in four conditions (ID8-2D, ID8-3D, ID8-KRAS-2D, and ID8-KRAS-3D) was analyzed by flow cytometry (d). The proportions of EdU-positive and GFP-positive cells were measured by flow cytometry (e). Experiments were repeated at least thrice. The values shown represent the mean ± SEM (* p < 0.05)