Extended Data Fig. 3: Effect of glycolysis inhibition in ISCs.
From: Metabolic adaptations direct cell fate during tissue regeneration
a, ATP rate assay in ISCs- and pAbs-enriched organoids showing the production of ATP through OXPHOS (MitoATP) or glycolysis (GlycoATP) in organoids. Each dot represents an independent experiment where five mice (n = 5) were pooled, and each lineage in each experiment was plated as 8 technical replicates. b, LC–MS was used to determine the steady-state level and fraction of glucose/ fructose 6-phosphate and 3-phosphoglycerate from 13C6-glucose. Glucose/ fructose 6-phosphate and 3-phosphoglycerate levels are shown as peak area in arbitrary units (a.u.) x fractional percentage of labelling with each tracer (n = 3/condition). Results are representative of 2 independent experiments. c, Experimental design for glycolysis inhibition in C57Bl/6 mice. d, H&E staining and immunofluorescence of ISCs (OLFM4+ cells) of control and 2-deoxyglucose (2-DG)-treated mice after one month of treatment. e, Quantification of mean OLFM4+ cells per crypt from (c). Each dot represents one mouse (n = 5). At least 30 crypts were analysed per mouse. f, Gene set enrichment analysis (GSEA) showing downregulation of the ISC gene signature in intestinal tissue from 2-DG–treated mice compared with control mice. g, Heat map illustrating transcription of genes in the ISC signature in intestinal tissue from control and 2-DG treated mice. Each row represents an individual mouse. This figure is adapted from our published patent (WO2024229094A1)50. Statistical significance: data represent mean ± s.e.m. Statistical significance was determined by two-tailed t-test in e. In each panel, asterisks indicate statistical significance (***P < 0.001).
