(A) A representative histogram showing reduced 2-NBDG uptake by AMPKαΔ/Δ bone marrow L-GMPs in vivo. (B) AMPK deletion significantly reduced glucose uptake of whole AML cells in the bone marrow and spleens, and L-GMPs in the bone marrow but not in the spleens (n=5). Immunoblotting (C) and immunostaining (D) assays revealed that AMPK deletion reduced the protein levels of Glut1, but not Glut4, in whole AML cells and L-GMPs, concomitant with increased Txnip1 levels. Scale bars indicate 5 μm. (E) Overexpression of Glut1 substantially rescued the defective clonogenicity of AMPKαΔ/Δ AML cells (n=4). (F) Immunoblotting against Glut1 using gene-edited AML cells revealed that Glut1 sgRNA clones 1 and 2 reduced Glut1 protein levels, whereas clone 3 had little effects. (G) Clonogenecity of the gene-edited cells as in (F). Glut1 sgRNA clones 1 and 2 significantly reduced clonogenicity, whereas clone 3 did not (n=4). (H) AML cells expressing sgRosa26 (n=5) or sgGlut1 (clone 1, n=5; clone 2, n=5) were transplanted. Deletion of Glut1 by sgRNAs significantly attenuated leukemogenesis. See also Figure S4.