Cell-viability assay

Exponentially growing cells (5 × 10³ cells per well) were plated in each well of a 96-well plate with 0.1 ml of RPMI 1640 + 10% (vol/vol) FCS containing the appropriate concentration of drug in triplicate, and cell viability was assessed after 48 hr as previously described (Smith et al., 2012). Log₁₀-fold selectivity heat maps for Figures 1C, 6A: briefly, IC50 values were calculated as shown in figures, then ratios of RTK/FLT3 (Figure 1C) or point mutant/BaF3 FLT3 ITD (Figure 6A) were calculated. Log₁₀-scale transformation of ratios, followed by conditional formatting using xcel spreadsheet software yielded the colored diagrams shown.