Figure 5.

Constitutively active Akt1 does not enhance viability or clonogenicity of p53-deficient FDM cells in the absence of IL-3. (a) Multiple independent clones of FDM cells of the indicated genotypes expressing iAkt1 or ieGFP were cultured in the presence of 4-OHT for 24 h before being starved of IL-3 over a 5-day time course. Cell viability was determined by PI exclusion using flow cytometry. (b) The same cells shown in (a) were removed from liquid culture and plated in medium containing 0.3% soft agar and 0.5 ng/ml IL-3. The numbers of colonies were counted after 10 days and expressed relative to the numbers of colonies from time 0 day. Results from (a) and (b) represent the mean and standard errors of three independent experiments. The numbers (n) of independent clones were as follows: WT iAkt1 n=5, WT ieGFP n=4, p53^−/− iAkt1 n=5, p53^−/− ieGFP n=5, FoxO3a^−/− iAkt1 n=5, FoxO3a^−/− ieGFP n=5