Figure 2. Functional effects of CITED1 misexpression in WiT49 cells.

(A) Relative to empty vector WiT49 control cells, overexpression of both CITED1ΔNES and wild-type CITED1 enhanced proliferation at each time point tested (*p >0.001), although the rate of change over time did not vary (p=0.29), and no significant difference was observed between CITED1ΔNES and wild-type CITED1. (B) When the three WiT49 cell lines were grown in soft agar (four biological replicates; open/closed circles and squares correspond to each experiment), CITED1ΔNES significantly enhanced colony formation relative to wild-type CITED1 (2.22 fold, p=0.0286) and to empty vector controls (5.01 fold, p=0.0286). Wild-type CITED1 colony count was not statistically different from empty vector control (p=0.2). (C) When xenotransplanted into immunodeficient mice, each WiT49 cell line showed similar growth characteristics as in soft agar. CITED1ΔNES tumors showed the highest mean tumor volumes relative to the other cell lines (p >0.0001; all data points shown). (D) Mean tumor weights were also different between the three cell lines (p=0.028) but showed greater variability. CITED1ΔNES weights were significantly greater than wild-type CITED1, although differences between wild-type and empty vector were not statistically different.