Enhancer landscapes defines transcriptional regulatory network of LSCs. (A) De novo motif analysis showing the enrichment of TFs at LSC enhancers compared with known motif background using HOMER. (B) STRING database analysis demonstrates that 11 of 12 LSC-specific transcription factor dependencies have putative protein–protein interactions. Red nodes indicate TFs previously coreported with leukemia in a literature search. (C,D) ChIP-qPCR of SPI1 and CEBPA binding at selected enhancers in LSCs. IgG enrichment was used as a negative control (n = 3; mean ± SD). (E) Colony formation assay at day 5 in LSCs infected with shRNAs targeting Spi1, Cebpa, and Runx1 (n = 3; mean ± SD). (F) Discovering transcription coregulators that share regulatory targets with SPI1, CEBPA, RUNX1, and MYB. (G) ChIP-qPCR of EP300 binding at selected enhancers in LSCs. IgG enrichment was used as a negative control (n = 3; mean ± SD). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. HOMER, hypergeometric optimization of motif enrichment; IgG, immunoglobulin G; qPCR, quantitative PCR.