Expression of ZNF198-FGFR1 activates both the AKT and ERK pathways, leading to phosphorylation at 14–3-3 binding sites of FOXO3a (T32) and BAD (S112), respectively. (A) (Left) Schematic diagram of ZNF198-FGFR1 fusion and truncation constructs. (Right) Expression of ZNF198-FGFR1 4ZF and 10ZF isoforms as well as activated truncation mutant PR/TK results in phosphorylation of FOXO3a (T32) and BAD (S112). Ba/F3 cells and cells expressing a kinase dead mutant 4ZF/ΔPR were included as negative controls. (B) Activation of the AKT and MAPK pathways in Ba/F3 cells stably expressing 4ZF and 10ZF isoforms resulted in phosphorylation of FOXO3a (T32) and BAD (S112), respectively. Cells stably expressing 4ZF were treated with U0126 and wortmannin at the indicated concentration for 90 minutes before immunoblotting. (C) Induced expression of 4ZF and 10ZF in TonBaF cells results in phosphorylation of FOXO3a (T32) and BAD (S112).