Extended Data Figure 4: The energy stress sensor LKB1–AMPK plays a pro-survival role and modulates glucose uptake and energy supply in pre-B ALL. | Nature

Extended Data Figure 4: The energy stress sensor LKB1–AMPK plays a pro-survival role and modulates glucose uptake and energy supply in pre-B ALL.

From: Metabolic gatekeeper function of B-lymphoid transcription factors

Extended Data Figure 4

a, Lkb1fl/fl mice were crossed with Cre-deleter strains for deletion at early pre-B-cell stages (Mb1) and in fully mature B cells (Cd21). B-cell populations in bone marrow and spleen (n = 3 litter mates) were characterized by flow cytometry analysis. b, The catalytic subunit of Ampk has two isoforms, α1 and α2. Analysis of published gene expression data (GSE38463)38 revealed that expression of the α1-form peaks at later stages of B-cell development, whereas expression of both Lkb1 and the α2-form of Ampk peaks in pre-B cells. For this reason, we studied the consequences of inducible ablation of Lkb1 and Ampka2 in mouse models of BCR–ABL1-transformed pre-B ALL cells. Protein levels of Lkb1 and Ampkα2 were verified by western blots. c, Viable cell counts upon Cre-mediated deletion of Lkb1 or Ampka2 or on treatment with an empty vector (EV). d, Apoptosis following Lkb1 deletion was monitored by annexin V/ 7-aminoactinomycin D (7AAD) staining. e, Colony-forming ability was assessed by serial re-plating upon deletion of Lkb1 in pre-B ALL cells. f, Glucose uptake and ATP levels (normalized to cell numbers) were measured following Cre-mediated deletion of Ampka2. g, Luciferase bioimaging of transplant recipient mice injected with Lkb1fl/fl pre-B ALL cells transduced with 4-OHT-inducible Cre or an empty vector and treated with tamoxifen (0.4 mg per mouse; n = 7 per group). h, Overall survival was assessed by a Kaplan–Meier analysis (P value calculated by Mantel–Cox log-rank test). i, Leukaemia samples developed in recipient mice (Fig. 2d) were genotyped for the presence of either floxed or deleted Lkb1 and Ampka2 alleles (n = 3 mice). Representative FACS plots and images from three independent experiments are shown (a, d, e). Data shown as mean ± s.d. from three independent experiments and assessed by two-way ANOVA (c) or two-tailed t-test (e, f). For gel source data, see Supplementary Fig. 1.

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