Fig. 2. High PDK activity defines +4 ISCs from EE lineage.

a Immunofluorescence of pPDH in small intestinal sections. pPDH⁺ cells are present in both crypts (arrows) and villi (arrowheads). On the right, magnification of intestinal crypts and villi containing pPDH⁺ cells. Scale bar, 100 μm. b Percentage of pPDH⁺ cells in every crypt position (62 pPDH⁺ cells form four mice were scored). c Double immunofluorescence for pPDH and BrdU. Representative image of four different mice is shown. Scale bar, 25 μm. d pPDH immunofluorescence on isolated crypts from mTert-GFP mice. Arrows mark pPDH+mTert+ cells. Asterisk marks non-specific fluorescence signal. Scale bar, 50 μm. e Quantification of pPDH⁺, mTert⁺, and pPDH⁺mTert⁺ cells in the intestinal crypts form three mTert-GFP mice. A total of 68 crypts were scored. f Double immunofluorescence for pPDH and ChgA on intestinal sections. Arrows indicate double positive cells, arrowheads indicate pPDH⁺ChgA⁻ cells. Scale bar, 50 μm. g Quantification of pPDH⁺, ChgA⁺, and pPDH⁺ChgA⁺ cells on intestinal sections from three mice (at least 50 crypts/mouse were scored). Data are presented as mean ± SEM. h Gene set enrichment analysis of positive regulators of glycolysis in Bmi1-GFP cells compared to Lgr5 ISCs from Lgr5-GFP and Lgr5-DTR mice (Jadhav et al.²⁰). i Heat map of the glycolytic gene signature from (a) showing the core enriched genes. Source data are provided as a Source Data file.