The FOXO3aS644A-GFP nuclear localization induces the expression of the FOXO3a target genes, p21 and Fas-L, inhibits proliferation, and induces apoptosis. (A) MV4-11 cells were infected as previously described in Figure 5A and were highly purified by flow cytometric cell sorting according to GFP expression after 48 hours of infection. p21^cip1 and Fas-L mRNA expression was quantified in triplicate in the purified cells by qRT-PCR, and their levels were expressed relative to HPRT mRNA levels. Results are expressed as a ratio to the control incubation with the vector alone. Vertical bars indicate SDs. (B) A total of 2.10⁴/mL MV4-11 cells was infected and highly purified as previously described in Figures 5A and 6A were incubated in triplicate in 10% FCS MEM and pulsed 6 hours with 1 μCi (37 kBq) [³H]thymidine. The amounts of radioactivity were determined after trichloracetic acid precipitation. Results are expressed as a ratio to the control condition. Results are obtained from 3 independent experiments in MV4-11 cells. The statistical significance was calculated versus the control condition (MV4-11 cells expressing the GFP protein alone [except where indicated by brackets]) by a Student t test. Vertical bars indicate SDs. (C) Infected MV4-11 cells were stained with annexin V–PE. Results are expressed as a percentage of annexin V–stained cells. Results were obtained from 6 independent experiments. The statistical significance was calculated versus the control condition (MV4-11 cells expressing the GFP protein alone [except where indicated by brackets]) by a Student t test. Vertical bars indicate SDs.