Skip to main content
. 2018 Apr 23;9(5):470. doi: 10.1038/s41419-018-0502-4

Fig. 5. The induction of Puma by FOXO3A depends on GSK-3.

Fig. 5

a Ba/F3 cells expressing CRISPR/Cas9 constructs targeting Luciferase or Foxo3a were infected with retrovirus encoding CRISPR/Cas9-resistant human FOXO3A control retrovirus (empty vector, EV). The cells were deprived of IL-3 in the presence or absence of CT98014 (CT, 0.75 µM) for 18 h and analyzed for apoptosis by Annexin V staining. The significance was tested by one-way ANOVA with post hoc Tukey’s multiple comparison test. Error bars represent 95% confidence intervals from four independent experiments (n = 4). b Ba/F3 expressing CRISPR/Cas9 constructs targeting Luciferase (crLUC) or a Foxo3a−/− single-cell clone (#4) were infected with retrovirus expressing human FOXO3A or none (empty vector, EV). The cells were deprived of IL-3 in presence or absence of CT98014 (0.75 µM, CT) for 18 h, then analyzed for apoptosis by Annexin V staining. Error bars represent SD from technical replicates. c Cells from b were harvested after 18 h and analyzed by western blotting with the antibodies indicated. d HCT116 p53−/− cells were transfected by Lipofectamine2000® with vectors encoding FOXO3A or control vector (empty). Twenty-four hours after transfection, the cells were treated with DMSO or GDC-0941 (10 µM) for 5 h. The cells were harvested and analyzed by western blotting, probing with antibodies as indicated.