American Society of Hematology

Figure 2

$Figure 2. Induced expression of a 14–3-3 antagonist R18 induces apoptosis in Ba/F3 cells transformed by ZNF198-FGFR1. (A) Schematic diagram of the structure and amino acid sequences of YFP-tagged R18 dimer and mutant. The R18 dimer construct contains 2 R18 motifs separated by a linker of 11 amino acids. (B) Inducible expression of YFP-tagged R18 dimer and mutant as well as stable expression of ZNF198-FGFR1 4ZF in the TonBaF cell lines. The GFP antibody applied cross-reacts with YFP. (C) TonBaF cells transformed by 4ZF are more sensitive to R18-induced apoptosis compared with control TonBaF-R18 cells. Cells were stained with PE-conjugated antiannexin V reagent and analyzed by FACS for the apoptotic population that is characterized as the fraction of annexin V/YFP double-positive cells in total YFP-positive cells (%). TonBaF cells inducibly expressing R18 cultured in the presence of IL-3 were included as controls. (D) R18 expression induced cleavage of PARP and caspase-3 in TonBaF R18 parental cells and 4ZF-transformed cells.$

Induced expression of a 14–3-3 antagonist R18 induces apoptosis in Ba/F3 cells transformed by ZNF198-FGFR1. (A) Schematic diagram of the structure and amino acid sequences of YFP-tagged R18 dimer and mutant. The R18 dimer construct contains 2 R18 motifs separated by a linker of 11 amino acids. (B) Inducible expression of YFP-tagged R18 dimer and mutant as well as stable expression of ZNF198-FGFR1 4ZF in the TonBaF cell lines. The GFP antibody applied cross-reacts with YFP. (C) TonBaF cells transformed by 4ZF are more sensitive to R18-induced apoptosis compared with control TonBaF-R18 cells. Cells were stained with PE-conjugated antiannexin V reagent and analyzed by FACS for the apoptotic population that is characterized as the fraction of annexin V/YFP double-positive cells in total YFP-positive cells (%). TonBaF cells inducibly expressing R18 cultured in the presence of IL-3 were included as controls. (D) R18 expression induced cleavage of PARP and caspase-3 in TonBaF R18 parental cells and 4ZF-transformed cells.

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