Figure 3: Spheroid IMCE-Ras cells demonstrated increased tumorigenicity both in vitro and in vivo. (A) Soft-agar analysis of monolayer and spheroid IMCE-Ras cells. Cells (5 × 103 cells per 35-mm well) were re-suspended in complete medium containing 0.35% agarose. After 14 days, colonies were methanol-fixed and stained with 0.005% crystal violet for visualization and counting. Triplicate experiments were performed. p < 0.05 by student’s t test. (B) Serial dilution of xenograft transplantation assay. Spheroid or monolayer cells at different dilutions were subcutaneously injected into both flanks of NOD/ SCID mice. N =4. p < 0.05 by student’s t test. The tumor formation and growth was monitored twice a week.