Differentially expressed proteins in LSC vs blast and LSC vs HSPC comparisons. (A) Heat map representation of protein expression of members of the LSC100 AML stem cell signature.9 (B) Log2 fold protein expression of proposed LSC markers in the indicated comparison. Each dot represents an individual AML sample. (C) Log2 fold-change (FC) in indicated comparison in FLT3-ITD/NPM1-mut AML (x-axis) vs FLT3-WT/NPM1-WT AML samples (y-axis). (D) Proteins significantly (padj < .01) overexpressed in both LSC vs blast and LSC vs HSPC comparisons in both AML subgroups. (E) CFU assay using MOLM13 cells after induced knockdown of MBOAT7 with 2 different hairpins (shMBOAT7 #1 and #2) and a nontargeting control (shNT). Each dot represents 1 culture, 2-sided Student t test. (F-G) Top 15 and bottom 15 proteins for LSC vs blast (left), LSC vs HSPC (middle), and combined (right) comparisons. (F) FLT3-ITD/NPM1-mut AML samples. (G) FLT3-WT/NPM1-WT cases. (H) Validation of selected LSC surface proteins by flow cytometry in n = 15 AML samples (n = 7 FLT3-ITD/NPM1-mut and n = 8 FLT3-WT/NPM1-WT cases) and n = 5 HSPC samples. (I) CFU assay using MOLM13 cells after induced knockdown of CRIP2 with 2 different hairpins (shCRIP2 #1 and #2) and a nontargeting control (shNT). Each dot represents 1 culture, 2-sided Student t test.