Fig. 1: Human CML LSCs have increased glucose metabolism compared to HSCs.

a Schematic overview of experimental design and omic dataset generation from human CML and non-leukaemic (healthy) samples. Created with BioRender.com (Agreement number: BL25IFA5C3). b Volcano plot of differentially expressed metabolism genes comparing CML LSCs and normal HSCs (CD34 + CD38−). Red coloured genes have a q value < 0.05 and blue coloured genes have q value < 0.1. The Benjamini-Hochberg adjustment was used to correct for multiple comparisons. c GSEA conducted using global Reactome pathway gene sets (see source data). d GSEA conducted using Hallmark gene sets. e CML-upregulated gene sets (from (d)) shown are glycolysis, oxidative phosphorylation and fatty acid oxidation. f Schematic overview of stable isotope labelling experiments. Tracers employed are ¹³C₆ glucose, ¹³C₅ glutamine and ¹³C₁₆ palmitate. g–i Volcano plots of metabolite fraction labelling following 24 h culture in ¹³C₆ glucose, (n = 4 each group; (g)), ¹³C₅ glutamine (n = 6 CML, n = 3 normal; (h)) and ¹³C₁₆ palmitate (n = 3 for each group; (i)), comparing CML CD34+ and normal CD34+ cells. Red coloured metabolites have q value < 0.05 and blue coloured metabolites have a q value < 0.1. Multiple unpaired t-tests were used and the two-stage step-up (Benjamini, Krieger, and Yekutieli) was used to correct for multiple comparisons. (LCMS raw data source: Kuntz et al.¹², for (g) and (i)). j Overlay of fold changes from (g) and (h) with transcriptomic data from (b). Significant changes in gene expression are indicated by gene symbol in blue. Significant changes in fraction labelling are indicated by metabolite name in red. GLUC (red bar) refers to fold change in glucose fraction labelling and GLUT (blue bar) refers to fold change in glutamine fraction labelling with average shown. * Refers to q value < 0.1. k Fractional labelling (n = 3 each normal or CML (and indicated subpopulations)) following 24 h culture in ¹³C₆ glucose. Mean and SEM are plotted and an one-way ANOVA was used with multiple comparisons corrected with the Benjamini-Hochberg adjustment. Source data are provided as a Source Data file.