Extended Data Fig. 7: Role of OGDH in intestinal homeostasis in vivo.
From: Metabolic adaptations direct cell fate during tissue regeneration
a, Experimental design. OGDH depletion in TRE-shOgdhCag-rtTA3 mice was induced in adulthood. Created in BioRender. Chaves-Perez, A. (2025) https://BioRender.com/1nvuf0r. b, qPCR analysis of Ogdh expression in TRE-shOgdhCag-rtTA3 and TRE-shRenCag-rtTA3 mice after 3 days on doxycycline. Each dot represents a different mouse (n ≥ 5). c, Kaplan–Meier survival curves of TRE-shOgdhCag-rtTA3 (n = 5) and TRE-shRenCag-rtTA3 (n = 6) mice under doxycycline treatment. d, Body weight relative to weight at baseline in TRE-shOgdhCag-rtTA3 (n = 5) and TRE-shRenCag-rtTA3 (n = 6) mice. e, Weight of food in the stomachs of TRE-shOgdhCag-rtTA3 and TRE-shRenCag-rtTA3 mice after 8 days on doxycycline. Each dot represents a different mouse (n ≥ 5). f, Experimental schematic for DM-αKG injections in adult C57Bl/6 mice. Created in BioRender. Chaves-Perez, A. (2025) https://BioRender.com/dimozin. g, Body weight relative to weight at baseline in mice treated with vehicle or DM-αKG at indicated doses. h, Kaplan–Meier analysis of survival of mice treated with DM-αKG at the indicated doses or with vehicle. i, H&E and immunofluorescence for BrdU, GFP and OGDH after 3 days on doxycycline. j, H&E and immunofluorescence for HNF4α on day 6 of doxycycline treatment. Dashed lines outline crypt structures. k,l, Quantification of BrdU (k) and Cl. Casp3 (l) levels in intestinal sections during doxycycline treatment of TRE-shRenCag-rtTA3 and TRE-shOgdhCag-rtTA3 mice. Each dot represents a crypt (n = 5 mice). m, H&E and immunofluorescence for β-catenin (Bcat), BrdU and HNF4α in mice that received 7 days of injections of DM-αKG (600 mg/kg). Dashed lines outline crypt structures. n, Representative smFISH image visualizing RNAs of various differentiation markers in intestinal crypts from TRE-shOgdhCag-rtTA3 and TRE-shRenCag-rtTA3. Dashed lines outline crypt structures and specific lineages in the intestinal epithelium. o, Immunofluorescence for BrdU, β-catenin and lysozyme (Lyz) after a 2-hour BrdU pulse in mice treated with DM-αKG-treated or vehicle. Dashed lines outline specific lineages within the crypts. p, Hyperplex immunofluorescence in colonic sections control and αKG-treated mice in steady-state conditions, showing ATOH1 (marking secretory cells), EphB2 (stem cells) and MUC2 (mature secretory cells). q, Quantification of ATOH1+/EphB2+ double-positive secretory progenitors from p. r, ABP staining in TRE-shOgdhCag-rtTA3 and TRE-shRenCag-rtTA3 mice. s, Immunofluorescence and quantification of OLFM4 intensity (arbitrary units) in TRE-shOgdhCag-rtTA3 and TRE-shRenCag-rtTA3 mice at day 4 of treatment, and in vehicle- and DM-αKG-treated mice at day 7 of treatment. Each dot represents one crypt from n ≥ 4 mice. Dashed lines outline crypt structures. This figure is adapted from our published patent (WO2024229094A1)50. Statistical analysis: Data represent mean ± s.e.m. Statistical significance was determined using a two-tailed t-test in b,e,q,s. Survival probabilities were estimated using the Kaplan–Meier method, and statistical differences between the survival curves were assessed using the log-rank (Mantel–Cox) test in c,h. Two-way ANOVA followed by Tukey’s HSD test was used in d,g. One-way ANOVA followed by Tukey’s HSD test was used in k, l. Asterisks denote statistical significance (*P < 0.05, **P < 0.01, ***P < 0.001). Schematics created with BioRender.com.
