Figure 3

hMDMX overexpression promotes anchorage-independent growth and tumor growth in vivo. A, B. Various VH10hTERT and HER cell lines were embedded in 0.3% agarose on a 0.6% agarose bottom-layer, with additional normal growth medium or growth medium containing 10 μM Nutlin-3 on top of the agarose. Colony outgrowth was monitored and pictures were taken 18 days (VH10) and 4 weeks (HER) after seeding. Representative pictures of several independent experiments are shown. C. Schedule for investigating in vivo growth of parental and transformed VH10 cells using the shell-less chicken CAM model. At embryonic development day (EDD) 7, 2.5 million cells were grafted onto a chicken CAM. Tumors were harvested at EDD17. D. Representative pictures of GFP-positive tumors, P-Histone-3 staining and vimentin staining. E. Tumor volumes of VH10hTERT control (N = 5), shRB-HRasV12-shp53 (N = 4) and shRB-HRasV12-HA-hMDMX (N = 5). F. Quantification of the number of mitotic (P-Histone-3 positive) cells. Statistical analysis was performed by one-way ANOVA followed by Bonferroni's Multiple Comparison Test. P < 0.05 was considered statistically significant.