Fig. 3: High glycolysis supports stem cell potential of pPDH⁺ intestinal epithelial cells.

a Immunofluorescence of pPDH and ChgA in small intestinal organoids. Notice that most cells are pPDH⁺ChgA⁺ (arrows). The experiment was repeated four times with similar results. Scale bar, 25 μm. b Immunofluorescence of pPDH and Ki67 in intestinal organoids. The experiment was done four times with similar results. Scale bar, 20 μm. c Single cells derived from intestinal organoids were infected with the Pdk1-mCherry reporter. Picture shows a 3-day growing organoid with few crypt cells positive for the reporter (representative image of 10 different experiments). Scale bar, 50 μm. d Immunofluorescence showing co-localization of the mCherry reporter and pPDH (representative image of three independent experiments is shown). Scale bar, 50 μm. e FACS-sorted mCherry⁻ and mCherry⁺ cells were plated in matrigel and cultured in ENR medium. Organoids were counted 7 days after plating (n = 3 independent sorting experiments). Data are presented as mean ± SD. f Intestinal crypts were grown in ENR medium in the absence of presence of DCA and organoids counted at day 7 (n = 6 technical replicates). A representative experiment is shown. The experiment was repeated three times with similar results. Data are presented as mean ± SD. Two tail t-test was used to determine statistical significance between groups. Scale bar, 750 μm. g H&E staining (top) and Ki67 IHC (bottom) of intestinal sections of mice treated or untreated with DCA 6 days post-IR (11 Gy). The graph shows the quantification of Ki67⁺ cells in the intestines of these mice (n = 2 mice per condition, at least 30 crypts/mouse were scored). Two tailed t-test (for comparisons between two groups) or one-way ANOVA (for comparisons of more than two groups) were used to determine statistical significance between groups (*p < 0.05; **p < 0.01; ***p < 0.001). Source data are provided as a Source Data file.