Downregulation of LSC genes in Myc^V394D-AML cells compared with Myc-AML cells. Myc/Cre^ERTMyc^fx/fx and Myc^V394D/Cre^ERTMyc^fx/fx AML cells were treated with 4-OHT to induce the deletion of endogenous Myc. (A-E) Myc- and Myc^V394D-AML cells were collected on day 4 of 4-OHT treatment and Myc deletion was confirmed. The differentially expressed genes between Myc- and Myc^V394D-AML cells are presented as a Venn diagram (A) and a volcano plot (B). The expression of LSC genes was analyzed by gene set enrichment analysis (C). The expression of selected LSC-related genes (D) and Myc-Miz1 target genes (E) were compared between Myc- and Myc^V394D-AML cells. (F) Kit⁺CD11b^low LSCs and Kit⁺CD11b^high leukemic blasts were purified from Myc-AML cells. The expression of selected LSC genes and Myc-Miz1 target genes was examined by quantitative real-time PCR (qRT-PCR) assay and compared. (G-H) Myc^V394D/Cre^ERTMyc^fx/fx AML cells were treated with 4-OHT to induce the deletion of endogenous Myc. Cells were collected on day 0 (vehicle), day 2, and day 4 after 4-OHT treatment. The expression of the selected genes was examined by qRT-PCR and normalized to messenger RNA (mRNA) levels for day 0 (G). The percentages of Kit⁺CD11b^low LSCs and Kit⁺CD11b^high leukemic blasts were examined by flow cytometry (H). *P < .05; **P < .01. fpkm, fragments per kilobase of transcript per million mapped reads.