Dimerization of MLL fusion proteins and FLT3 activation synergize to induce multiple-lineage leukemogenesis
J. Clin. Invest. Ryoichi Ono, et al. 115:919 doi:10.1172/JCI22725 [
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Figure 4Synergistic transformation of murine hemato-poietic progenitors by MLL fusion genes and FLT3-ITD in vitro. (
A) Schematic representation of the retroviral constructions expressing FLT3-ITD. (
B) Transformation assay of the cells immortalized by MLL-SEPT6, after transduction with FLT3-ITD in the pMY-IRES-EGFP construct shown in
A in the presence (+) or absence (–) of IL-3. (
C) Western blot analysis of proteins extracted from PlatE cells transfected with the constructs shown in
A and each vector alone as a control, after immunoprecipitation using the anti-FLT3 Ab (lanes 1–4). Each lysate was blotted with the anti-FLT3 Ab. Lane 1, pMY-IRES-EGFP alone; lane 2, pMY-FLT3-ITD-IRES-EGFP; lane 3, pMYpuro alone; lane 4, pMYpuro-FLT3-ITD. (
D) Myeloid immortalization assay using the pMYpuro constructs shown in
A. The bar graph shows numbers of colonies obtained after each round of replating in methylcellulose (average ± SD). (
E) Myeloid transformation assay using the sequential transduction with FLT3-ITD or control (pMYpuro alone) after MLL-SEPT6, MLL-ENLs, or mock (pMXs-neo alone) in the presence (+) or absence (–) of IL-3.
