A critical role of NF-κB signaling in the myeloid transformation induced by AML1 mutants. (A) Schematic presentation of structures of the AML1 mutants. Runt indicates Runt domain, and TAD indicates transactivating domain. (B-C) Nuclear translocation assays of p65 in HEK293T cells transduced with AML1 mutants as indicated, 20 minutes after the addition of TNF-α. AML1 (B) and S291fsX300 (C). Scale bar represents 10 μm. (D-E) In vitro kinase assays of IKKα (D) or IKKβ (E) in HEK293T cells transduced with AML1 or S291fsX300, 20 minutes after the addition of TNF-α. Kinase activities were detected by autoradiography and quantified with ImageJ Version 1.41o software.31 (F-G) Nuclear translocation assays of p65 in HEK293T cells transduced with AML1 mutants as indicated, 20 minutes after the addition of TNF-α. A224fsX228 (F) and R293X (G). Scale bar represents 10 μm. (H) Colony counts from the serial replating assays of AML1-mutants-transformed cells with BMS-345541. Error bars show mean ± SEM.
