LSC-enriched population expresses self-renewal gene-expression signature. Primary leukemia cells harvested from spleens were sorted into immunophenotypic subpopulations (Mac-1^High, Mac-1^LowKit^–Sca-1^–, Mac-1^LowKit⁺Sca-1^–, and Mac-1^LowKit⁺Sca-1⁺). (A) Sorted cells were plated in methylcellulose CFAs. The number of colonies that developed per 4000 cells plated is noted. Numbers represent the average of 7 independent experiments. 95% confidence intervals are indicated. (B) 10⁵ sorted cells were intravenously injected into SCID/Beige mice. Disease-free survival is indicated. Each group consisted of 7 or 8 mice. P values noted reflect comparisons between the (*) Mac1^LowKit⁺Sca1⁺ vs Mac1^High groups and (**) Mac1^LowKit⁺Sca1⁺ and unsorted groups. (C-E) Cells from each sorted subpopulation were submitted for RNA sequencing. (C) The NRAS^G12D-directed gene-expression signature specific to self-renewing HSCs⁹  is enriched in the LSC-enriched subpopulation (Mac-1^LowKit⁺Sca-1⁺). (D) The MLL-AF9-mediated self-renewal gene-expression signature is enriched in the LSC-enriched subpopulation (Mac-1^LowKit⁺Sca-1⁺). (E) The NRAS^G12V leukemia self-renewal signature was derived by identifying the NRAS^G12V-mediated gene-expression profile that is also present in the LSC-enriched subpopulation.