Disruption of palmitoylation promotes leukemia cell growth and leukemia progression in vivo. (A) Examination of palmitoylation status of endogenous FLT3-ITD by using the APE analysis in different MV4;11 cell clones that were either not edited (ITD) or were edited for C563S heterozygous (ITD/CS Het) or homozygous (ITD/CS Homo) mutations via CRISPR/Cas9. (B) Quantification of surface FLT3 levels in ITD, ITD/CS Het, and ITD/CS Homo MV4;11 clones determined by flow cytometry. (C) Examination of downstream signaling in individual MV4;11 clones using WB analysis. (D) Relative cell growth of ITD, ITD/CS Het, and ITD/CS Homo MV4;11 clones at different days, as determined by enumeration of the cells. (E) Different MV4;11 clones were plated in methylcellulose media and the number of colony-forming leukemia cells quantified after 7 to 10 days. (F) Bioluminescence imaging of NSG mouse recipients of xenografts of unedited FLT3-ITD or of ITD/CS Homo–edited MV4;11 clones expressing firefly luciferase-T2A-mCherry at 1 and 2 weeks. (G) Quantification of bioluminescence signals of mouse recipients of xenografts as shown in panel F. Each symbol represents an individual mouse (ITD, n = 7; ITD/CS Homo, n = 8). Means ± standard error of the mean are presented as vertical lines. (B,D-E) Data analyses were performed in triplicate; results are presented as means ± standard deviation. In all relevant panels, *P < .05; **P < .01; ***P < .001; ns, not significant, as determined by 2-tailed Student t test.