Figure 1. Establishment of FLT3-ITD and TKD mutant transfectants and drug screening.

(A) Schematic primary structure of FLT3-ITD isolated from MV4-11 cells. The positions of N676 and F691 lie on tyrosine kinase domain (TKD) 1, and those of D835 and Y842 are on TKD2. TM, transmembrane domain; JM, juxtamembrane domain; ITD, internal tandem duplication. (B) Cells were treated with increasing concentrations of quizartinib, midostaurin, or gilteritinib (1–256 nM) for 4 days, and cell viabilities were determined by WST-8 assay. The viabilities of drug-treated cells relative to those of untreated cells were calculated and are shown here as the mean ± SD from three independent experiments. (C and D) Cells were treated with or without 10 nM quizartinib (C), gilteritinib, or midostaurin (D) for 6 h, and immunoblotting using the indicated antibodies was performed. (E) Cells were treated with 10–1000 nM concentrations of various inhibitors, listed in Supplementary Table 1, for 4 days, and cell viabilities were determined by WST-8 assay. The clustered image map was created at the CIMminor site using the average IC₅₀ values from triplicated experiments, as described in the Methods section.